JP7209309B2 - Degassing, heating, and high-pressure processing methods for food - Google Patents

Description

The present invention relates to a method for degassing, heating, and high-pressure treatment of food.

In Patent Document 1, as a method for producing a seasoning, ingredients such as sardines and raw oysters containing or added with proteolytic enzymes are maintained in a temperature range of 40 ° C. to 60 ° C. under a pressure of 50 MPa to 100 MPa. , discloses a method for shortening the ripening period of seasonings by promoting the action of enzymes while inhibiting the growth of microorganisms.

In Patent Document 2, as a method for producing processed fish products such as turnip sushi, a relatively low pressure of 100 MPa or less is applied to fish meat materials such as yellowtail, thereby destroying the tissue of the fish meat materials and seasoning such as salting. Techniques for shortening the time required have been disclosed.

Non-Patent Document 1 describes a method for injecting functional saccharides (5% trehalose and 5% cyclodextrin) into wild plants using an impregnation apparatus sealed at a pressure of 30 kg/cm ² .

In Non-Patent Documents 2 and 3, light pickling of vegetables (eggplants, cucumbers, etc.) is produced in a short time by applying the impregnation method used when injecting a hardening agent into wood to lightly pickling foods. The content to be performed is described.

Japanese Patent No. 3475328 JP 2013-55912 A

The 4th Trehalose Symposium (November 8, 2000) Nihon Agricultural Newspaper (issued on October 3, 1989) Hokkoku Shimbun (published September 28, 1989)

High-pressure treatment makes it possible to impregnate the interior of food with liquid seasonings and enzymes in a short period of time. can be expected.
However, when high-pressure treatment is used in combination with conventional food manufacturing methods, there is a balance with the conditions of the conventional method, and even if the same manufacturing method is used, the ingredients contained in each individual food are different. Equivalent effects cannot be expected under different conditions, and there are optimal treatment conditions for each food product and manufacturing method that applies high-pressure treatment.

In the case of "fruits pickled in syrup" such as apples, plums and apricots, there is a demand to increase the sterilization effect while maintaining the texture and to extend the storage period.

In order to solve the above problems, the degassing, heating, and high-pressure treatment method applied to the production of food according to the present invention is that the food is Japanese pears or grapes pickled in syrup, and the Japanese pears or grapes are Put it in a container such as a bag and deaerate, hold only Japanese pears or grapes and syrup in the container, and put the Japanese pears or grapes in a container such as a polyethylene bag at 80 MPa or more and 300 MPa or less, Then, it is treated at 40° C. or higher and 70° C. or lower for 15 to 60 minutes.

According to the degassing/heating/high-pressure treatment method according to the present invention, umami (taste component) components such as free amino acids and sugars can be increased in a short period of time. , the manufacturing period, which used to take one to one and a half years, can be shortened to several days.

Moreover, when the method of the present invention is applied to the production of syrup-pickled fruits such as apples, plums and apricots, the sterilization effect can be enhanced and the storage period can be extended.

In addition, when pickled livestock meat is produced by the method of the present invention, the texture (softness) of the meat is improved in addition to the above, and when pickled tofu is produced by the method of the present invention, it can be crushed with the tongue. It is possible to obtain a cream cheese-like texture with a hardness of 1, and to increase free amino acids. The increase was made possible even in boiling water.

Furthermore, when seafood lees pickles and soft smoked products are produced by the method of the present invention, not only the effect of the high pressure heating treatment but also the synergistic effect of the sterilization effect of the alcohol and smoke contained in the sake lees causes the growth of bacteria. etc. is suppressed.

A diagram showing the components detected in Narazuke when the lees bed is treated at 70°C or higher and when treated with the conventional treatment method. A diagram comparing the conventional manufacturing method and the degassing/heating/high-pressure treatment method of the present invention. Photographs of narazuke of eggplant, narazuke of Japanese radish, narazuke of gourd processed by the method of the present invention Diagram explaining the sterilization principle of degassing, heating, and high-pressure treatment Photograph of lightly pickled vegetables for people with difficulty chewing produced by degassing, heating, and high-pressure treatment according to the method of the present invention A diagram explaining the process of producing raw soy sauce pickles of vegetables produced by degassing, heating, and high pressure treatment according to the method of the present invention. Graph showing the amount of penetration of raw soy sauce into radish Graph showing the penetration amount of raw soy sauce for various vegetables Photograph showing the effect of soy sauce permeation into Japanese radish Graph comparing the effect of soy sauce permeation into Japanese radish Photos and graphs showing the effect of cutting radishes Graph showing the relationship between apple syrup concentration and self-destructive germination sterilization effect Graph showing results of spontaneous germination-inducing bactericidal effect when acidulant (phytic acid) was added to aqueous sucralose solution A graph comparing the sugar content and acidity of fruit juice and syrup after subjecting five types of apples to degassing, heating, and high-pressure processing. Photograph showing the number of bacteria in apples pickled in syrup Photograph showing the number of bacteria in apples pickled in syrup Photograph showing the number of bacteria in apples pickled in syrup A graph comparing the sugar content of fruit juice and syrup and the physical properties of the flesh of apricots subjected to degassing, heating, and high-pressure treatment under four types of treatment conditions. A graph comparing the acidity of extracts obtained by combining apricots, sake, and sugar and subjecting them to degassing, heating, and high-pressure treatment Graph showing the results of cyanide analysis of apricots and plums pickled in degassed, heated, and high-pressure syrup A diagram showing an example of an apricot pickled in syrup A diagram showing an example of a method for pickling plums in syrup Photograph of plum syrup preserved at 5°C for 4 months Photograph of plum syrup preserved at 25°C for 4 months Photograph comparing the shape of ume with holes and ume without holes after soaking in syrup Photograph of syrup-pickled Japanese pears produced by the same method as plum syrup-pickled A method for producing plum wine made from plums pickled in syrup, which is made by combining plums and sugar, degassing, heating, and high-pressure processing. Comparison graph of component content (acidity) of umeshu by high-pressure processing time Comparison graph of ingredient content (polyphenol) in umeshu by high-pressure treatment time Comparison graph of component content (sucrose decomposition rate) of umeshu by high-pressure treatment time Graph comparing the conventional method, the method of the present invention, and the commercial product for the acidity and polyphenols of plum wine Comparison graph of sucrose decomposition rate (indicator of maturation) of umeshu by manufacturing method Comparison graph of color tone of plum wine by manufacturing method Graph showing the concentration of free amino acids in examples 1 to 3 of miso-pickled pork Graph showing glucose concentration of examples 1 to 3 of miso-pickled pork Graph showing the difference in hardness after heating depending on the heating temperature of miso-pickled pork Graph showing the difference in ease of biting after heating depending on the heating temperature of miso-pickled pork Graph showing the difference in the ease of biting off after heating due to the difference in processing time of miso-pickled pork Graph showing changes in ease of chewing after heating due to differences in seasoning liquid for miso-pickled pork Graph showing acid-soluble nitrogen content of Beef Miso-zuke Examples 1-3 Graph showing glucose concentration of Beef Miso-zuke Examples 1-3 Graph showing the difference in hardness (breaking stress) after heating of examples 1 to 3 of beef miso pickles Graph showing the concentration of free amino acids in tofu lees pickles Graph showing hardness of tofu lees pickles (examples 1 to 3) Graphs showing the hardness of tofu lees pickles, miso pickles, and shio-koji pickles (Examples 3 to 5) Diagram explaining the formation and decomposition of guanylic acid Graph showing guanylic acid contained in each treated shiitake mushroom Graph showing the salt content in each processed shiitake mushroom Graph showing hardness (breaking stress) of shiitake mushrooms with each treatment Graph showing the salt concentration of radish pickled in sake lees Graph showing ease of biting off daikon radish pickled in sake lees Graph showing the degree of discoloration of radish pickled in sake lees (a) is a photograph of radish before high pressure heat treatment, and (b) is a photograph of radish after high pressure heat treatment. Graph showing the salt concentration of Nakashima greens pickled in sake lees Graph showing the hardness of Nakashimana stems pickled in sake lees Graph showing the degree of discoloration of Nakashima greens pickled in sake lees (a) is a photograph of Nakashimana before high pressure heat treatment, (b) is a photograph of Nakashimana after high pressure heat treatment Graph showing the salt concentration of daikon radish pickled in soy sauce Graph showing ease of biting off soy sauce pickled daikon radish (a) is a photograph of radish before high pressure heat treatment, and (b) is a photograph of radish after high pressure heat treatment. Graph showing degree of cell destruction of soy sauce-pickled daikon radish by treatment temperature Graph showing the amount of umami components in soy sauce-pickled radishes Graph showing the salt concentration of Nakashimana pickled in soy sauce Graph showing the firmness of the stems of Nakashimana pickled in soy sauce Graph showing the degree of discoloration of Nakashimana pickled in soy sauce (a) is a photograph of Nakashimana before high pressure heat treatment, (b) is a photograph of Nakashimana after high pressure heat treatment Graph showing the sugar content of plums soaked in syrup Graph showing acidity of plums pickled in syrup Graph showing the hardness of plums soaked in syrup Graph showing the degree of discoloration of plums soaked in syrup (a) is a photograph of plums before high-pressure heat treatment, and (b) is a photograph of plums after high-pressure heat treatment. Graph showing the sugar content of pears pickled in syrup Graph showing the firmness of pears soaked in syrup Graph showing the degree of discoloration of pears soaked in syrup (a) is a photograph of pears before high-pressure heat treatment, and (b) is a photograph of pears after high-pressure heat treatment. Graph showing the degree of cell destruction of pears soaked in syrup by treatment temperature Graph showing the sugar content of grapes soaked in syrup Graph showing hardness and ease of chewing of grapes soaked in syrup Graph showing the degree of discoloration of grape pulp soaked in syrup (a) is a photograph of grapes before high pressure heat treatment, (b) is a photograph of grapes after high pressure heat treatment Graph showing sugar concentration of miso marinated beef thigh Graph showing the hardness of miso-marinated beef thigh after heating (a) is a photograph of beef thigh before high pressure heat treatment, and (b) is a photograph of beef thigh after high pressure heat treatment. Graph showing amino acid content of miso marinated beef thigh Graph showing the sugar concentration of miso-pickled beef tongue Graph showing the hardness of miso-pickled beef tongue (a) is a photograph of beef tongue before high-pressure heat treatment, and (b) is a photograph of beef tongue after high-pressure heat treatment. Graph showing amino acid content of miso-pickled beef tongue Graph showing the glucose concentration of beef thigh meat pickled in koji Graph showing hardness after heating of koji-pickled beef thigh (a) is a photograph of beef thigh before high pressure heat treatment, and (b) is a photograph of beef thigh after high pressure heat treatment. Graph showing amino acid content of beef thigh meat pickled in koji Graph showing glucose concentration of koji-pickled beef tongue Graph showing hardness of koji-pickled beef tongue after heating (a) is a photograph of beef tongue before high-pressure heat treatment, and (b) is a photograph of beef tongue after high-pressure heat treatment. Graph showing amino acid content of koji-pickled beef tongue Graph showing the glucose concentration of hard tofu pickled in sake lees Graph showing the hardness of hard tofu pickled in sake lees Graph showing the degree of discoloration of hard tofu pickled in sake lees (a) is a photograph of hard tofu before high pressure heat treatment, and (b) is a photograph of hard tofu after high pressure heat treatment. Graph showing amino acid content of hard tofu pickled in sake lees Graph showing the glucose concentration of Fukuragagi pickled in sake lees Graph showing the hardness of Fukuragagi pickled in sake lees Graph showing the degree of discoloration of Fukuragagi pickled in sake lees (a) is a photograph of Fukuraguri before high-pressure heat treatment, and (b) is a photograph of Fukurugi after high-pressure heat treatment. Graph showing the glucose concentration of squid pickled in rice lees Graph showing the hardness of squid pickled in sake lees Graph showing the degree of discoloration of squid pickled in sake lees (a) is a photograph of squid before high-pressure heat treatment, and (b) is a photograph of squid after high-pressure heat treatment. Graph showing glucose concentration in soft-smoked Fukurage Graph showing the firmness of soft-smoked fukuragi Graph showing the degree of discoloration of soft-smoked Fukuragi (a) is a photograph of Fukuraguri before high-pressure heat treatment, and (b) is a photograph of Fukurugi after high-pressure heat treatment. Graph showing glucose concentration in soft-smoked squid Graph showing firmness of soft-smoked squid Graph showing degree of discoloration of soft-smoked squid (a) is a photograph of squid before high-pressure heat treatment, and (b) is a photograph of squid after high-pressure heat treatment.

The best mode for carrying out the present invention is described below as ``narazuke of vegetables'', ``lightly pickled vegetables'', ``raw soy sauce pickled vegetables'', ``fruits pickled in syrup'', ``pickled meat'', and ``tofu. pickles,” “processed mushroom products,” additional experiments on these, and production of pickled fish and shellfish in sake lees and soft smoke.

Narazuke of vegetables 1. Experimental method 1-1 Sample preparation Prepare eggplant, radish, and gourd as vegetables, remove the air inside the plant tissue by sun drying and degassing, and promote the penetration of the components of the lees bed. Especially for eggplants with a smooth surface, the permeation of the components of the lees bed was accelerated by making many holes with a kenzan or by boiling them once in boiling water and cooling them (blanching treatment).

1-2 Processing The eggplants were coated with a lees bed and placed in a polyethylene bag, degassed, and then placed in the polyethylene bag and subjected to heat and high pressure treatment under conditions of 100 MPa, 70° C., and 60 hours.
The radish was coated with a lees bed and placed in a polyethylene bag, then degassed. While placed in the polyethylene bag, it was subjected to heating and high pressure treatment under the conditions of 100 MPa, 70° C., and 18 hours.
For cucumbers, we used the lees bed that was produced in a short period of time by heat treatment. After applying this dregs and placing it in a polyethylene bag, it was degassed, and then, while it was placed in a polyethylene bag, it was subjected to heat and high pressure treatment under the conditions of 100 MPa, 50° C., and 60 hours.

In order to produce the lees bed in a short period of time, 10% sugar and 5% salt were added and treated at a high temperature of 70° C. or higher for several hours to several days to accelerate the Maillard reaction. It takes a long time to prepare ordinary lees bed, but according to the method of this embodiment, a forced lees bed having a unique flavor of Narazuke lees bed can be produced in several hours to several days.
When the forged lees bed is used, GC-MS analysis detects specific components (the components enclosed in the frame in FIG. 1), making it easy to distinguish from the conventional product.

In addition, the following (Table 1) shows the sterilization effect of Bacillus spores when the dregs are heated and high pressure treated, and from this table, Bacillus spores can no longer be detected under the conditions of 100 MPa, 65 ° C. or higher, and 60 hours. I understand.

2. Test results and discussion Fig. 2 is a diagram comparing the conventional method and the heat and high pressure treatment method of the present invention. In the method of the present invention, ginjo lees containing salt and mirin are salted in one step. After pickling in sake lees containing salt and mirin, it was pickled in ginjo lees. In both cases, the production period (one year to one and a half years) of the traditional method could be greatly shortened.

Fig. 3 is a photograph of the eggplant, radish and gourd produced by the above heat and high pressure treatment, and all narazuke were comparable in taste, aroma and texture to narazuke produced by conventional traditional methods.

The following (Table 2) shows the measurement results of the number of viable bacteria and the number of yeasts in Narazuke of eggplant produced by heat and high pressure treatment. From this table, Narazuke produced by the method of the present invention is substantially sterile. I understand.

FIG. 4 is a diagram illustrating the principle of sterilization by heating and high-pressure treatment. At room temperature, only a portion of spore-forming bacteria germinate, but with heating, all spore-forming bacteria germinate. Once germinated bacteria do not stop on the way, and the germinated bacteria are killed by heat treatment at 50° C. or higher.
Conventional normal temperature and pressure treatment requires final sterilization at high temperature, but in the case of heat and high pressure treatment, high temperature treatment is not required, so it is possible to prevent food quality deterioration that accompanies sterilization. .

3. Effect of treatment at 100 MPa or more When the vegetable narazuke manufacturing method described in paragraph (0025) and after was examined by changing only the pressure condition of the high pressure treatment condition in the range of 100 to 300 MPa, the results shown in Table 3 were obtained. It was confirmed that the invention is also effective in the pressure range of 100-300 MPa.

Lightly Pickled Vegetables for People with Difficulties in Chewing 1. Experimental Method 1-1 Preparation of Samples Carrots, eggplants and Japanese radishes were prepared and cut to a thickness of about 1 cm.

1-2 Processing Treatment The above cut vegetables were placed in a polyethylene bag together with a seasoning liquid (commercially available lightly pickled base) and pectinase, degassed, and placed in the polyethylene bag. Heating and high-pressure treatment were performed under conditions of minutes.
After that, each vegetable was subjected to normal pressure heating at 0.1 MPa for 3 to 15 hours. Specifically, carrots were treated for 15 hours, eggplants for 3 hours, and radishes for 3 hours and 5 hours.
After that, the enzyme was deactivated by heating in boiling water for 5 minutes.

2. Test Results and Discussion FIG. 5 is a photograph of lightly pickled vegetables produced by the heat and high pressure treatment according to the method of the present invention.
As shown in Table 4, all of the vegetables shown in FIG. 5 were extremely soft enough to melt in the mouth so that even those with difficulty chewing could eat them.

3. Effect of treatment at 100 MPa or more When the method for producing lightly pickled vegetables described in paragraph (0037) and after was examined by changing only the pressure condition of the high pressure treatment condition in the range of 100 to 300 MPa, the results shown in Table 4 were obtained. It was confirmed that the present invention is effective even in the pressure range of 100-300 MPa.

Vegetables Pickled in Raw Soy Sauce 1. Experimental Method 1-1 Preparation of Samples Carrots, Japanese radishes, cucumbers, eggplants, burdocks, Chinese yam, and lotus roots were prepared and cut into 0.5-1 cm thick pieces.
1-2 Processing As shown in FIG. 6, the above vegetables cut into 0.5-1 cm thickness are dried in the sun. This process removes moisture, making it easier for the seasoning liquid (raw soy sauce) to permeate.
Next, the above-mentioned cut vegetables were placed in a polyethylene bag together with a seasoning liquid (raw soy sauce), degassed, and degassed and heated under the conditions of 100 MPa, 50° C., and 20 minutes while being placed in the polyethylene bag.・High pressure treatment is applied.
Figure 7 shows the color tone and salt content of the sun-dried radish cut into 0.5 cm thickness and the sun-dried radish cut into 1 cm thickness, immediately after the treatment and the next day, when the conditions of degassing, heating, and high pressure treatment are changed. It is a graph which shows the measured result. From this graph, it can be seen that the method of the present invention allows the seasoning liquid (raw soy sauce) to easily permeate the radish.
FIG. 8 is a graph showing the results of infiltration of seasoning liquid (raw soy sauce) into various vegetables including the radish by degassing, heating, and high-pressure treatment. From this graph, it can be seen that the method of the present invention is effective for other than radish.

2. Test results and discussion As shown in Figures 6, 7 and 8, the color tone and salt content of the seasoning liquid permeated the vegetables uniformly, and the production period was greatly shortened from about two weeks to 20 minutes. . Moreover, as shown in Table 5, a bactericidal effect was confirmed, and long-term storage was possible.

3. Comparative Examples by Different Shapes of Radish In addition to the above, the permeation of the seasoning liquid was investigated, especially the radish.
As a material, Fukuhomare produced by Ishikawa Agricultural Research Institute was cut into round slices with a thickness of 3 cm. 20%, 40%, 60%, 80% and 100%, and the heat/high pressure treatment was 100 MPa, 30° C. or 50° C. for 24 hours.

FIG. 9 is a photograph showing the effect of soy sauce permeation into radish, and FIG. 10 is a graph comparing the effect of soy sauce permeation into radish. It can be seen that increasing the concentration of the liquid equalizes the permeation effect inside and outside. However, it is not suitable as a pickled food due to its high salt concentration.

FIG. 11 is a photograph and a graph showing the effect of making cuts in the radish. By making cuts in FIG. I understand. However, even with this method, the liquid seasoning did not sufficiently permeate into the interior.

4. Effect of treatment at 100 MPa or more When the method for pickling vegetables in raw soy sauce described in paragraph (0038) and after was examined by changing only the pressure condition of the high pressure treatment condition in the range of 100 to 300 MPa, the results are shown in Table 5. , confirmed that the present invention is effective even in the pressure range of 100 to 300 MPa.

Syrup Pickling of Fruits 1. Experimental Method 1-1 Preparation of Samples As fruits, apples (Shinano Gold, etc.), apricots (Shinzanmaru, etc.), and plums (Ishikawa No. 1, etc.) were prepared.

1-2 Processing Apples, apricots, and plums were each placed in a polyethylene bag together with syrup, deaerated, and placed in the polyethylene bag under the conditions of 100 MPa, 65°C to 75°C, and 30 minutes to 60 minutes. It was subjected to heat and high pressure treatment.

2. Test results and discussion 2-1 Self-defeating germicidal effect 10 ⁷ to 10 ⁸ CFU/g of Bacillus subtilis spores were inoculated into the material pickled in apple syrup, and the self-defeating germicidal effect was examined after each treatment. The results are shown in FIG.
From FIG. 12, it can be seen that the degassing/heating/high pressure treatment has a higher sterilizing effect than the normal temperature/normal pressure treatment or the heating/normal pressure treatment, and specifically has a sterilizing effect of about 2 log (CFU/g). In particular, the lower the syrup concentration, the higher the bactericidal effect.

FIG. 13 is a graph showing the results of the spontaneous germination-inducing bactericidal effect when an aqueous solution of sucralose, which is a high-intensity sweetener, is used as a substitute for syrup, and an acidulant (phytic acid) is added thereto. , It can be seen that the sterilization effect is greatly enhanced by using a low-concentration sweetener with an added acidulant and degassing, heating, and high-pressure treatment.

2-2. Degassing, Heating, and High-Pressure Treatment of Apples Five types of apples were prepared: Tsugaru, Shuuei, Shinanosweet, Shinanogold, and Fuji.
The above five types were subjected to degassing, heating and high pressure treatment at 100 MPa, 65° C. for 30 minutes, and the sugar content and acidity of fruit juice and syrup were compared. The results are shown in FIG.
From FIG. 14, it can be seen that the higher the temperature of the heating/high-pressure treatment, the closer the sugar content of the fruit juice and the syrup, and that the acidity is affected by the variety.

Table 6 is a graph showing the results of bacteria detection when the production temperature and storage period of apple degassing, heating, and high pressure treatment are changed. indicates that the storage period is 4 months. From these tables, storage at room temperature for 3 months, preferably 2 months, is considered safe.

In addition, FIG. 15 is a photograph showing the state of bacteria on the “fruit” side in the case of 4 months in Table 6 (manufacturing temperature 65 ° C., storage temperature 25 ° C.), and FIG. , storage temperature 25 ° C.), and FIG. It is a photograph which shows the state of bacteria.

2-3. Hardness improvement (retention)
Calcium lactate was added to Fuji and Shinano Gold to improve hardness, and good results were obtained.

2-4. Preservability Table 9 is a table showing the detection results of microorganisms when apples subjected to heat and high pressure treatment according to the method of the present invention were preserved at 5°C and room temperature (25°C).

From Table 9, it can be seen that microorganisms do not propagate for nearly 6 months when stored at 5°C. In addition, even if it is stored at room temperature, it is ensured for 3 months. Moreover, it can be seen from Table 9 that the apples stored at 5°C show almost no change in appearance even after 6 months.

2-5. Degassing/heating/high-pressure treatment for apricots After degassing and packaging apricots and 40% syrup, they were subjected to degassing/heating/high pressure treatment at 100 MPa at 65-75°C for 30-60 minutes. As a result, as shown in FIG. 18, syrup pickles with raw texture and raw flavor were obtained.
The frozen apricots and white liquor were combined and subjected to degassing, heating, and high-pressure treatment at 100 MPa, 65° C., and 30 to 120 minutes. As a result, as shown in FIG. 19, an apricot-flavored liqueur was obtained due to the high organic acid extraction effect.
Table 10 shows the bactericidal effect of apricots (cultivar: Heiwa) pickled in syrup. From this table, it can be seen that the degassing/heating/high-pressure treatment method of the present invention has a sterilization effect equivalent to that of the autoclave treatment.

Table 11 shows data on the number of bacteria in apricot syrup soaked for 0 months, and Table 12 shows the results of the storage test.

Degassed, heated, and high-pressure-treated apricots and plums pickled in syrup were analyzed for cyanide. As a result, as shown in FIG. 20, cyanide in the pulp was hardly increased compared to the unprocessed raw material.
Moreover, FIG. 21 is a photograph of pickled apricots in syrup.

2-6. Plum pickled in syrup Fig. 22 is a diagram showing an example of plum pickled in syrup. The part is hollowed out, followed by blanching water exposure, followed by heating and high pressure treatment. The conditions for the heating/high pressure treatment are 100 MPa, 65° C., and 30 minutes. After that, it was allowed to stand at normal pressure at 50° C. for 40 hours to make the flavors of the fruit and syrup uniform.

Table 13 is a table showing the results of a storage test of raw plums not subjected to heat and high pressure treatment, and Table 14 is a table showing the results of a storage test (0 months) of plums soaked in syrup that were subjected to heat and high pressure treatment. Table 15 is a table showing the results (one month) of storage tests of plums soaked in syrup that were subjected to heat and high pressure treatment, and Table 16 is the results of storage tests of plums soaked in syrup that were subjected to heat and high pressure treatment. (2 months), Table 17 is a table showing the results of a storage test (3 months) of plums soaked in syrup subjected to heat and high pressure treatment, and Table 18 is a table showing syrup subjected to heat and high pressure treatment. It is a table|surface which shows the result (4 months) of the preservation|save test of the pickled plum.

FIG. 23 is a photograph of plum syrup-preserved at 5° C. for 4 months, and FIG. 24 is a photograph of plum syrup-preserved at 25° C. for 4 months.

FIG. 25 is a photograph showing a comparison of the shapes of plums that have been punched and those that have not been punched after being pickled in syrup.

2-5. The conditions for heating and high-pressure treatment of Japanese pears in syrup are 100 MPa, 65° C., and 30 minutes. After that, it was allowed to stand at normal pressure at 50° C. for 40 hours to make the flavors of the fruit and syrup uniform.
FIG. 26 is a photograph of Japanese pears pickled in syrup produced by applying the above production method. Furthermore, as shown in Table 19, the sterilization effect of Japanese pears soaked in syrup according to the present invention was improved.

3. Effect of treatment at 100 MPa or more When the fruit syrup production method described in paragraph (0049) and after was examined by changing only the pressure condition of the high pressure treatment condition in the range of 100 to 300 MPa, the results shown in Table 19 were obtained. It was confirmed that the invention is also effective in the pressure range of 100-300 MPa.

Plum wine While the above has described pickling plums in syrup, the present invention can also be applied to the production of plum wine.
That is, conventionally, three ingredients of plum, sugar, and sake are mixed, treated at normal temperature and pressure for 72 hours (3 days), and then aged for about 3 months. As shown in Figure 27, ume and sugar are put in a pack, deaerated, heated, and subjected to high pressure treatment (100 MPa, 50°C, 72 hours) to extract an extract, which is mixed with sake to make umeshu. .
The ratio of ume and sugar and the treatment conditions are, for example, 100 to 300 g of sugar per 100 g of ume (fruit pulp), pressure of 100 MPa or more, heating temperature of 35 to 65° C., and high pressure treatment time of 24 to 72 hours. .
As a result, as shown in Figures 28-1, 28-2, 28-3, 28-4, 29, and 30, the conventional method (72 hours) was sufficient for plum extract (acid, polyphenol). However, it can be extracted considerably by the method of the present invention (72 hours). Similarly, decomposition of sucrose, which is a measure of maturation, hardly occurs in the conventional method, but it progresses considerably in the medium-high pressure method. Furthermore, the color (b* value, index of brown color) is darker in the medium-high pressure method than in the conventional method. It can be seen that when the method of the present invention is aged for 1 month, the color depth is almost the same as that of the conventional method aged for 3 months or longer.
The reason why the ripening of umeshu proceeds quickly by the method of the present invention is that extracts (acids, polyphenols) and enzymes that promote ripening (sugar-degrading enzymes (invertase)) are rapidly extracted from ume plums, and are processed under heat and high pressure. This is because the decomposition of sugar proceeds quickly.

3. Effect of treatment at 100 MPa or more When the method for producing plum wine described in paragraph (0083) and after was examined by changing only the pressure condition of the high-pressure treatment condition in the range of 100 to 300 MPa, the results shown in Table 20 were obtained. is effective even in the pressure range of 100 to 300 MPa.

pickled meat
1. Pork pickles 1-1 Experimental method 1-1-a Preparation of samples Prior to processing, the seasoning liquid shown in Table 21 below is applied to both sides of the pork loin sliced to a thickness of about 1 cm, and 2/3 weight of the meat is applied. It was placed in a polyethylene bag and deaerated and packaged with a deaeration degree of 99% using a deaeration packaging machine.

Here, the thickness of the pork is 1 cm and the amount of the seasoning liquid is 2/3 of the weight of the meat, but the present invention is not limited to the thickness of the pork and the amount of the seasoning liquid. Any degree of vacuum may be used as long as the air in the bag is sufficiently removed.

1-1-b Processing The deaerated packaged sample was processed as shown in Table 22 below. In the conventional method, miso liquid was used as the seasoning liquid and stored in a refrigerator (normal pressure, 4° C.) for 72 hours. In Examples 1 to 3, miso liquid was used as the seasoning liquid, and heat/high pressure treatment at 100 MPa was performed at treatment temperatures of 35, 45, and 55° C. for 16 hours.
In Example 4, the seasoning liquid was heated at 100°C for 10 minutes to inactivate the enzymes, and a heated miso paste was used.
In Example 5, miso liquid was used as the seasoning liquid, and heat treatment at 100 MPa and high pressure was performed at a treatment temperature of 45° C. for 1 hour.
In Example 6, sake lees liquid was used as the seasoning liquid, and heat treatment at 100 MPa and high pressure was performed at a treatment temperature of 45° C. for 16 hours.
In Example 7, salt koji was used as the seasoning liquid, and heat treatment at 100 MPa and high pressure was performed at a treatment temperature of 45° C. for 16 hours. The heating and high pressure treatment was performed using "Marugoto Extract TFS-20" manufactured by Toyo Koatsu Co., Ltd.

1-1-C Measurement of Taste Components Samples processed by the conventional method and Examples 1 to 3 were used to measure free amino acid concentration and glucose concentration as taste components.
Specifically, after homogenizing the meat after each treatment, 5 ml of an 8% trichloroacetic acid aqueous solution was added to 5 g of the sample, vigorously shaken, filtered, and the clarified liquid was brought to a constant volume of 50 ml. The total concentration of 20 major amino acids contained in the constant-volume liquid was measured and converted to the concentration of free amino acids contained in the original sample. Amino acids were measured using a Hitachi high-speed amino acid analyzer L-8900. Further, the glucose concentration of the constant-volume liquid was measured using a glucose CII test manufactured by Wako Pure Chemical Industries, Ltd., and converted to the glucose concentration contained in the original sample. Three replicates were performed per treatment, and the mean and standard deviation were determined.

1-1-d Texture Analysis Using the samples processed by the conventional method and Examples 1 to 7, "hardness" and "easy to bite off" were measured as indicators of texture.
Specifically, after removing the seasoning liquid from each processed meat and wiping off moisture on the surface of the meat with a paper towel, the sample was placed in a new plastic bag and degassed and packaged (vacuum degree 99%).
After the packaged sample was boiled in hot water at 70° C. for 1 hour, it was cooled with running water for 30 minutes, and the meat was cut into approximately 1 cm squares using a razor. Using Rheometer CR-500DX manufactured by Sun Science Co., Ltd., the cut sample was compressed by 9 mm in the direction perpendicular to the muscle fiber on the side without a razor blade, and the stress when compressed by 5 mm was measured. As an index of "hardness", the stress (shear stress) at the time of cutting was used as an index of "easy to bite off".
Twelve pieces of 1 cm square were measured for each sample, and the average value of 10 pieces excluding the maximum and minimum values was used as the measured value of the sample. The above operation was repeated 3 times per treatment, and the average value and standard deviation were obtained.

1-2. Test Results/Discussion 1-2-a Free Amino Acid Concentration FIG.
In the conventional method and Examples 1 to 3, the free amino acid concentration is higher than that of the raw meat before treatment. higher than the law.
Among Examples 1 to 3, Example 3, which was treated at the highest temperature, had the highest concentration of free amino acids. Therefore, in Examples 1 to 3 using heat and high pressure treatment, the umami component can be increased in a shorter time than the conventional method, and the treatment temperature of 55° C. is considered to increase the umami component the most.

1-2-b Glucose Concentration FIG. 32 shows the glucose concentrations of the samples processed by the conventional method and Examples 1-3.
In the conventional method and Examples 1 to 3, the glucose concentration is higher than the raw meat before treatment, and in Examples 1 to 3, the glucose concentration is higher than the conventional method, although the treatment time is significantly shorter than the conventional method. It was about the same. Further, among Examples 1 to 3, Example 3 with the highest treatment temperature had the highest glucose concentration. Therefore, in Examples 1 to 3 using heat and high pressure treatment, it is considered that the glucose concentration in the seasoning liquid can be increased in a shorter time than the conventional method.

1-2-c Change in Hardness after Heating Depending on Treatment Temperature FIG. 33 shows the hardness after heating of the samples processed by the conventional method and Examples 1-4.
The hardness of the conventional method was almost the same as that of no treatment, but Examples 1 to 3 were softer than those of no treatment. Further, among Examples 1 to 3, Examples 2 and 3 were the softest, and the hardness was about half that of the non-treated ones. Furthermore, in Example 4, in which the miso enzyme was deactivated by heating, although the same processing treatment as in Example 2 was performed, the hardness was almost the same as that of no treatment. Therefore, the fact that the meat became tender after heating in Examples 1 to 3 is considered to be due to the synergistic effect of the enzymes contained in the seasoning liquid and the processing treatment (heating and high pressure treatment).

1-2-d Changes in easiness of biting off after heating depending on treatment temperature Fig. 34 shows the easiness of biting off after heating of the samples processed by the conventional method and examples 1 to 4.
The conventional method and examples 2 and 3 are easier to bite off than no treatment, and examples 2 and 3 are easier to bite off than the conventional method despite the treatment time being significantly shorter than the conventional method. was becoming On the other hand, since Example 1 was almost the same as no treatment, it is considered necessary to treat at a temperature higher than 35 ° C in order to make it easier to bite off the meat after heating using heat and high pressure treatment. . In addition, since there was no significant difference between the untreated meat and the ease of biting off in Example 4, it is considered that an enzyme contained in the seasoning liquid is necessary to make the meat after heating easy to bite through.

1-2-e Changes in ease of biting off after heating due to differences in treatment time FIG.
Example 5, which was subjected to heating and high pressure treatment at 100 MPa and 45° C. for 1 hour, was inferior to the conventional method, but was easier to bite off than no treatment. Therefore, in the case of the treatment conditions of 100 MPa and 45.degree.

1-2-f Changes in ease of chewing after heating due to differences in seasoning liquid FIG.
Both Example 6 using sake lees liquid as the seasoning liquid and Example 7 using shio-koji (salted rice malt) were almost as easy to bite off as Example 2 using miso liquid as the seasoning liquid. Therefore, the present invention is not limited to pickling in miso, but is considered to be a technology that can be applied to general foods such as pickling in sake lees and pickling in shio-koji, in which livestock meat is pickled in a seasoning solution containing enzymes.

1-2-g Bactericidal effect As shown in Table 24, the miso-pickled pork performed in the present invention had an improved bactericidal effect.

2. Beef pickles 2-1 Experimental method 2-1-a Preparation of samples Prior to processing, a miso seasoning liquid (78% miso, 11% sugar, 11% mirin %) was applied to 2/3 of the weight of the meat, placed in a polyethylene bag, and degassed and packaged with a degassing degree of 99% using a degassing packaging machine.

Here, the thickness of the beef is 1 cm and the amount of the seasoning liquid is 2/3 of the weight of the livestock meat, but the present invention is not limited to the thickness of the beef and the amount of the seasoning liquid. Any degree of vacuum may be used as long as the air in the bag is sufficiently removed.

2-1-b Processing Treatment The processing shown in Table 23 below was performed on the vacuum-packaged sample. In the conventional method, miso liquid was used as the seasoning liquid and stored in a refrigerator (normal pressure, 4° C.) for 72 hours. In Examples 1 to 3, heat treatment at 100 MPa and high pressure were performed at a treatment temperature of 45° C. for 4, 8 and 16 hours, respectively.
The heating and high pressure treatment was performed using "Marugoto Extract TFS-20" manufactured by Toyo Koatsu Co., Ltd.

2-1-c Measurement of Taste Components Samples processed by the conventional method and Examples 1 to 3 were used to measure free amino acid concentration and glucose concentration as taste components.
Specifically, after homogenizing the meat after each treatment, 5 ml of an 8% trichloroacetic acid aqueous solution was added to 5 g of the sample, vigorously shaken, filtered, and the clarified liquid was brought to a constant volume of 50 ml. As an indicator of protein degradation products (free amino acids and peptides), the nitrogen content in the trichloroacetic acid extract was measured by the Kjeldahl method. Further, the glucose concentration of the constant-volume liquid was measured using a glucose CII test manufactured by Wako Pure Chemical Industries, Ltd., and converted to the glucose concentration contained in the original sample. Three replicates were performed per treatment, and the mean and standard deviation were determined.

2-1-d Texture Analysis Using the samples processed by the conventional method and Examples 1 to 3, "hardness" and "easy to bite off" were measured as indicators of texture.
Specifically, after removing the seasoning liquid from each processed meat and wiping off moisture on the surface of the meat with a paper towel, the sample was placed in a new plastic bag and degassed and packaged (vacuum degree 99%).
After the packaged sample was boiled in hot water at 70° C. for 1 hour, it was cooled with running water for 30 minutes, and the meat was cut into approximately 1 cm squares using a razor. Using Rheometer CR-500DX manufactured by Sun Science Co., Ltd., the cut sample was compressed by 9 mm in the direction perpendicular to the muscle fiber on the side without a razor blade, and the stress when compressed by 5 mm was measured. As an index of "hardness", the stress (shear stress) at the time of cutting was used as an index of "easy to bite off".
Twelve pieces of 1 cm square were measured for each sample, and the average value of 10 pieces excluding the maximum and minimum values was used as the measured value of the sample. The above operation was repeated 3 times per treatment, and the average value and standard deviation were obtained.

2-2. Test Results/Discussion 2-2-a Acid-Soluble Nitrogen (Free Amino Acids and Peptides) Concentration FIG.
In the conventional method and Examples 1 to 3, the acid-soluble nitrogen concentration is higher than the raw meat before treatment, and in Examples 1 to 3, the acid-soluble nitrogen concentration is was higher than the conventional method.
Further, among Examples 1 to 3, Example 3 with the longest treatment time had the highest acid-soluble nitrogen concentration. Therefore, in Examples 1 to 3 using heat and high pressure treatment, the umami component can be increased in a shorter time than the conventional method, and the treatment time of 16 hours is considered to increase the umami component the most.

2-2-b Glucose Concentration FIG. 38 shows the glucose concentrations of the samples processed by the conventional method and Examples 1-3.
In the conventional method and Examples 2 and 3, the glucose concentration is higher than the raw meat before treatment, and in Examples 2 and 3, despite the treatment time being significantly shorter than the conventional method, the glucose concentration is higher than the conventional method. It was about the same. Further, among Examples 1 to 3, the glucose concentration was the highest in Example 3, which had the highest treatment temperature and had a treatment time of 16 hours. Therefore, in Examples 2 and 3 using heat and high pressure treatment, it is considered that the glucose concentration in the seasoning liquid can be increased in a shorter time than the conventional method.

2-2-c Change in Hardness after Heating Depending on Treatment Temperature FIG. 39 shows the hardness after heating of the samples processed by the conventional method and Examples 1-3.
The hardness of the conventional method was almost the same as that of no treatment, but Examples 1 to 3 were softer than those of no treatment. In addition, among Examples 1 to 3, Example 3 was the softest and had a hardness of about 2/3 of the untreated.
Therefore, in the case of the treatment conditions of 100 MPa and 45° C., it is thought that heating and high pressure treatment should be performed for 16 hours in order to make the meat easier to bite off after heating.

2-2-d Bactericidal effect As shown in Table 24, the beef miso marinated according to the present invention had an improved bactericidal effect.

3. Effect of treatment at 100 MPa or more When the method for producing pickled livestock meat described in paragraph (0086) and after was examined by changing only the pressure condition of the high-pressure treatment condition in the range of 100 to 300 MPa, the results shown in Table 24 were obtained. It was confirmed that the invention is also effective in the pressure range of 100-300 MPa.

Tofu pickles 1. Experimental method 1-1 Sample preparation Prior to processing, Ishikawa prefecture's specialty hard tofu was sliced into 1 cm or 5 cm thick slices and wrapped in gauze. The seasoning liquid shown in 1 was applied to 1/2 weight of hard tofu, placed in a polyethylene bag, and degassed and packaged at a vacuum degree of 99% using a degassing packaging machine.
Here, the thickness of the hard tofu is 1 cm or 5 cm, and the amount of the seasoning liquid is 1/2 the weight of the hard tofu, but the present invention is not limited to the thickness of the hard tofu and the amount of the seasoning liquid. Also, the degree of vacuum of the degassed packaging may be any degree of vacuum as long as the air inside the bag is sufficiently removed.

1-2 Processing The processing shown in Table 26 below was performed on the deaerated and packaged sample. In the conventional method, hard tofu was sliced into 5 cm thick slices and stored in a refrigerator (normal pressure, 4° C.) for 9 months using sake lees liquid as a seasoning liquid.

In Example 1, hard tofu was sliced into 1 cm thick slices, and a sake lees liquid was used as the seasoning liquid, and the slices were subjected to heating and high pressure treatment at 100 MPa and 45° C. for 90 hours.
In Example 2, hard tofu was sliced into 5 cm thick slices, and sake lees liquid was used as the seasoning liquid. After pretreatment, the slices were stored in a refrigerator (normal pressure, 4°C) for 1 week, and then heated at 100 MPa and 45°C under high pressure. Treatment was carried out for 16 hours.

Pretreatment is a process to suppress variations in the surface and internal components of hard tofu, and by storing it in cold storage before heating and high pressure treatment, the ingredients and enzymes of the seasoning liquid are soaked into the inside of the tofu in advance. go to Here, pretreatment was performed using degassed packaged samples, but a large amount of tofu was soaked in the seasoning liquid at once using a barrel or tray, stored in a refrigerator, and then taken out and degassed individually. Heating and high-pressure treatment may be performed after packaging.

In Example 3, hard tofu was sliced into 1 cm thick slices in order to improve the penetration of the seasoning liquid, and the slices were subjected to heating and high pressure treatment at 100 MPa and 45° C. for 16 hours using sake lees liquid as the seasoning liquid.
In Example 4, hard tofu was sliced into 1 cm thick slices, and a miso paste liquid was used as the seasoning liquid, followed by heating and high pressure treatment at 100 MPa and 45° C. for 16 hours.
In Example 5, hard tofu was sliced into a thickness of 1 cm and subjected to heating and high pressure treatment at 100 MPa and 45° C. for 16 hours using salt koji liquid as the seasoning liquid. The heating and high pressure treatment was performed using "Marugoto Extract TFS-20" manufactured by Toyo Koatsu Co., Ltd.

1-3 Measurement of Taste Component Samples processed by the conventional method and Examples 1 to 3 were used to measure free amino acid concentration as a taste component. Specifically, the seasoning liquid was removed together with the gauze from the sample after each treatment, and 1 cm of the surface layer of the tofu was cut off and sampled separately as the “surface layer” and the remaining portion as the “inside”.
Examples 3-5 were sampled entirely as the surface layer. After separately homogenizing the sampled surface layer and the inside, 5 ml of an 8% trichloroacetic acid aqueous solution was added to 5 g of the sample, vigorously shaken, filtered, and the clarified liquid was brought to a constant volume of 50 ml. The total concentration of 20 major amino acids contained in the constant-volume liquid was measured and converted to the concentration of free amino acids contained in the original sample. Amino acids were measured using a Hitachi high-speed amino acid analyzer L-8900. Only one repetition was performed for the conventional method, and three repetitions were performed for each of Examples 1 to 3, and average values and standard deviations were determined.

1-4 Food Texture Analysis Using the samples processed by the conventional method and Examples 1 to 5, the hardness of the sample surface was measured and used as a measure of the food texture.
Specifically, the seasoning liquid was removed together with the gauze from the sample after each treatment, and 1 cm of the surface layer of the sample was cut off. The cut sample is further cut into 1 cm squares, and using a rheometer (CR-500DX manufactured by Sun Science Co., Ltd.), a cylindrical plunger with a diameter of 10 mm is used to compress the hard tofu by 9 mm in the direction from the surface to the inside, The maximum stress was used as an index of hardness. Ten pieces of 1 cm square were measured per sample, and the average value was taken as the measured value of the sample. The above operation was repeated once in the conventional method and three times in each of Examples 1 to 5, and the average value and standard deviation were obtained.

2 Test Results and Discussion 2-1 Free Amino Acid Concentration FIG.
In the conventional method, after refrigerating for 1 week, the surface layer is about 1000 mg/100 g, and the inside is about 900 mg/100 g. After refrigeration, the concentration of free amino acids increased with the length of the refrigeration storage period.

On the other hand, in Example 1, the surface layer is about 2500 mg/100 g and the inside is about 1600 mg/100 g, and although the treatment time is as short as 90 hours, both the surface layer and the inside have more free amino acids than those stored in a refrigerator for one month. increased concentration.
In Example 2, the surface layer was about 1500 mg/100 g and the inside was about 1000 mg/100 g, and the surface layer was almost the same as the one after refrigerating for one month and the inside after one week.
Example 3 was about 1500 mg/100 g, which was almost the same as the one month refrigerated storage despite the short treatment time of 16 hours.
From the above results, by using heat and high pressure treatment, it is possible to achieve a free amino acid concentration equivalent to that of products manufactured by conventional methods over a month in a few days (16 to 90 hours) of processing. Conceivable.

2-2 Difference in Hardness Depending on Treatment Method FIG. 41 shows the hardness of the samples treated by the conventional method and Examples 1 to 3.
In the conventional method, the product after refrigerating for one week was slightly harder than before treatment. It is considered that this is because the osmotic pressure of the seasoning liquid removes water from the hard tofu and tightens the structure.
The one after refrigerating for one month was slightly softer than before the treatment, but the difference was such that it could hardly be distinguished by sensory evaluation.
After refrigerating for 9 months, it was much softer than before treatment, and had a hardness that could be crushed with the tongue (1×10 ⁴ N/m ² or less), and when eaten, had a cream cheese-like texture. was

On the other hand, in Example 1, although the treatment time was as short as 90 hours, it was as hard as the product stored in the refrigerator for 9 months by the conventional method, and it was so hard that it could be crushed with the tongue, and it tasted like cream cheese. It had a nice texture.
Examples 2 and 3 are slightly harder than Example 1, but are hard enough to be crushed with gums (5×10 ⁴ N/m ² or less), and when eaten, are clearly softer than before treatment. , It had a cheese-like texture.

2-3 Verification of seasoning liquid Fig. 42 shows the hardness of samples processed in Examples 3-5.
Both Example 4 using miso liquid as the seasoning liquid and Example 5 using shio-koji had approximately the same hardness as Example 3 using sake lees liquid as the seasoning liquid.
Therefore, the present invention is not limited to pickling in sake lees, but is considered to be a technique that can be applied to general foods such as miso pickling and shio-koji pickling, in which tofu is pickled in a seasoning solution containing enzymes.

2-4 Sterilization effect As shown in Table 27, the pickled tofu prepared in the present invention has an improved sterilization effect.

3. Effect of treatment at 100 MPa or more When the method for producing pickled tofu described in paragraph (0112) and after was examined by changing only the pressure condition of the high-pressure treatment condition in the range of 100 to 300 MPa, the results shown in Table 27 were obtained. It was confirmed that the invention is also effective in the pressure range of 100-300 MPa.

Processed mushroom product 1. Experimental method 1-1 Sample preparation Prior to processing, raw shiitake mushrooms from which the stems were removed and a seasoning liquid (mixed with soy sauce and butter at a ratio of 1:2) of 30% by weight of the raw shiitake mushrooms were mixed. It was placed in a polyethylene bag and degassed and packaged at a degree of vacuum of 99% using a degassing packaging machine.
Here, the whole shiitake mushroom cap portion is used, and the amount of the seasoning liquid is 30% of the shiitake mushroom, but the present invention is not limited to the shape of the shiitake mushroom and the amount of the seasoning liquid. Any degree of vacuum may be used as long as the air inside the bag is sufficiently removed.

1-2 Processing The processing shown in Table 28 below was performed on the degassed and packaged samples.
In conventional method 1, heat treatment was performed at 0.1 MPa and 98° C. for 10 minutes as boiling sterilization.
In conventional method 2, heat treatment was performed at 0.2 MPa and 121° C. for 20 minutes as retort sterilization.
In conventional method 3, heat treatment was performed at 0.1 MPa and 70° C. for 30 minutes as heat sterilization.
In the examples using this patent, heat treatment was performed at 100 MPa and 70° C. for 30 minutes as heat/high pressure sterilization. The heating and high pressure treatment was performed using "Marugoto Extract TFS-20" manufactured by Toyo Koatsu Co., Ltd.

1-3 Measurement of Guanylic Acid Using the samples processed by Conventional Methods 1 to 3 and Examples, the concentration of guanylic acid, which is the umami component of mushrooms, was measured. Specifically, the method was as follows.
After wiping off the seasoning liquid from the sample after each treatment, distilled water of the same weight as the sample was added and homogenized, followed by centrifugation (3000 rpm, 10 minutes) to recover the supernatant. The supernatant was filtered through a filter with a pore size of 0.45 μm and analyzed by high performance liquid chromatography. High performance liquid chromatography was performed under the conditions shown in Table 29 below. Guanylic acid (guanosine monophosphate) manufactured by Wako Pure Chemical Industries, Ltd. was used as a standard.

1-4 Salt Content Analysis Salt content was measured as a taste component using the samples processed by Conventional Methods 1 to 3 and Examples. Specifically, the amount of sodium contained in the supernatant for guanylic acid measurement was measured by atomic absorption spectrometry and converted to the concentration of sodium chloride contained in the original sample.

1.5 Texture analysis Using the samples processed by conventional methods 1 to 3 and examples, the hardness of the cap portion was measured by the method of Kasuga et al. was measured using That is, using a rheometer (CR-500DX manufactured by Sun Science Co., Ltd.), a cylindrical plunger with a diameter of 1 mm was used to compress a point 5 mm away from the axis of the cap part from the direction of the folds, and when it penetrated The stress was obtained. Measurements were taken at four points per sample, and the average value was taken as the measured value of the sample. Five replicates were performed per treatment, and the mean and standard deviation were determined.

2 Test results and discussion 2-1 Guanylic acid generation mechanism Guanylic acid is a typical umami substance contained in mushrooms. FIG. 43 shows a schematic diagram of the production and decomposition of guanylic acid.
Guanylic acid is produced by degrading nucleic acid (RNA) contained in the nucleus of cells with an enzyme (ribonuclease). Therefore, when mushroom cells are damaged, nucleic acid present in the cell nucleus is easily degraded by ribonuclease present in the cytoplasm, and guanylate is readily produced.
Since the cells of dried shiitake mushrooms are damaged by drying, the umami component guanylic acid is more likely to increase than fresh shiitake mushrooms, and has been used as an ingredient for dashi since ancient times. Guanylic acid is converted to tasteless guanosine when degraded by phosphatases. A ribonuclease that produces guanylate has high activity at 65-70°C, and a phosphatase that degrades guanylate has high activity at 40-60°C. Furthermore, since phosphatase is inactivated at 65°C or higher, it is known that shiitake mushrooms are most likely to develop their umami when heated at 65-70°C.

2-2 Guanylic Acid Concentration FIG. 44 shows the guanylic acid concentrations of the samples processed by Conventional Methods 1 to 3 and Example.
Raw shiitake mushrooms before treatment contained almost no guanylic acid, but the guanylic acid concentration increased by heating. In the example, the concentration of guanylic acid increased more than three times that of conventional methods 1 and 2, and about 1.5 times that of conventional method 3.
From the above results, it is considered that the guanylic acid contained in shiitake mushrooms can be increased by using the method of the present invention as compared with the usual heat sterilization method. The reason for the increase in guanylic acid is thought to be the possibility that the ribonuclease activity was enhanced by the heat/high pressure treatment, and the possibility that the nuclear membrane was damaged by the heat/high pressure treatment.

2-3 Salinity Fig. 45 shows the salinity of the samples processed by Conventional Methods 1 to 3 and Example.
The salinity increased depending on the heating time but not on the heating method. In addition, since shiitake mushrooms have many voids in the tissue, it is thought that the seasoning liquid permeated due to air-liquid replacement when degassed and packaged.

2-4 Texture Fig. 46 shows the breaking ability of the samples processed according to Conventional Methods 1 to 3 and Example.
From this figure, no significant difference in texture was observed.

2-5 Bactericidal Effect As shown in Table 30, the mushroom processed product of the present invention has improved bactericidal effect.

3. Effect of treatment at 100 MPa or more When the manufacturing method of the processed mushroom product described in paragraph (0129) and after was examined by changing only the pressure condition of the high pressure treatment condition in the range of 100 to 300 MPa, the results shown in Table 30 were obtained. It was confirmed that the invention is also effective in the pressure range of 100-300 MPa.

Additional experiment (vegetables)
(1) Pickled radish in sake lees / processing conditions Pressure: 1000 atmospheres (100 MPa)
Temperature: 40-70°C
Time: 1 to 5 hours Production details Vegetables: Daikon radish (blue neck)
Pickles: seasoning lees (sake lees 70%, salt 10%, sugar 20%)
Production method: Peeled daikon radish was cut into a half-moon shape with a width of 1 cm.
- Evaluation method Salinity: each sample was pulverized and filtered, and the filtrate was used as a measurement sample and measured with a salinity meter (manufactured by Toa Denpa Co., Ltd.). As shown in FIG. 47, the high-pressure heating treatment permeates the components and increases the salt concentration. Differences in salt concentration due to treatment temperature and treatment time tend to be slightly proportional.
Ease of biting off: each sample was prepared into a square cube of 1 cm high and 2 cm wide, and the stress when it was penetrated to 6 mm by a 5 mmφ cylindrical plunger was measured with a rheometer. As for the measurement results, as shown in FIG. 48, the daikon lees-pickled daikon radish that has been subjected to high-pressure heating improves the easiness of biting off in proportion to the treatment temperature and treatment time. Furthermore, its ease of biting off is close to that of ready-made products.
Color tone: L*, a*, and b* were measured by the reflection method using a color difference meter (manufactured by Nippon Denshoku Co., Ltd.) for the color tone of the sample under each processing condition. As shown in FIG. 49, the color of the sake lees permeates into the radish by high-pressure heating. It is believed that the penetration is roughly proportional to the treatment temperature and treatment time. Also, FIG. 50(a) shows the color tone before the high pressure heating treatment, and FIG. 50(b) shows the color tone before the high pressure heating treatment.
Microbial hygiene test method: Suspend each sample in physiological saline, serially dilute, and add each diluted sample to standard agar medium (Eiken, for general viable cell count) and 0.01 (w/v) % chloramphenicol. Potato dextrose agar medium (Eiken, for fungi) was smeared (in the case of standard agar, the pour method was used). After culturing at 30 to 37° C. for 3 days, the number of microbial colonies formed was counted, and the number of viable general bacteria and fungi per 1 g of sample was calculated. With reference to the standards of the Food Sanitation Act, the standard values were 300 cfu/g or less for the general viable count and 1000 cfu/g or less for the fungal count. Non-detection was defined as 50 cfu/g or less for both general viable bacteria and fungi.
(Table 31A) shows the results of microbial hygiene inspection by treatment temperature (all treatment times are 3 hours), and (Table 31B) shows the results of microbial hygiene inspection by treatment time (all treatment temperatures are 70°C).

In the table, above the standard means 100,000/g or more for general viable bacteria, 1000/g or more for fungi, and below the standard means 100,000/g or less for general viable bacteria, 1000/g or less for fungi, and no Detection refers to the limit of detection, 50/g or less.
As shown in the table, the number of viable bacteria is below the sanitary standard value for both treatment temperature and treatment time. This is thought to be due not only to the effects of high-pressure heating, but also to the synergistic effect of the sterilizing effect of the alcohol contained in the sake lees soaked into the daikon radish, which inhibited the growth of bacteria.

(2) Nakashima greens pickled in sake lees / processing conditions Pressure: 1000 atmospheres (100 MPa)
Temperature: 40-70°C
Time: 15-45 minutes Production details Vegetables: Nakashima greens Pickled bed: seasoning lees (sake lees 50%, sake 20%, salt 10%, sugar 20%)
Production method: The ratio of stems and leaves of Nakashima greens was set to 1:1, and the mixture was placed in a bag together with pickles of the same weight as the total weight and vacuum-packaged.
- Evaluation method Salinity: each sample was pulverized and filtered, and the filtrate was used as a measurement sample and measured with a salinity meter (manufactured by Toa Denpa Co., Ltd.). As shown in FIG. 51, the high-pressure heating treatment permeates the components and increases the salt concentration. The treatment temperature increases up to 60°C in proportion to the increase in salinity, but it decreases at 70°C. The stem cut with a jar was placed on its side, and the stress when it penetrated to 10 mm through the center was measured with a rheometer. As shown in FIG. 52, the measurement results show that the stalks of Nakashimana pickled in sake lees that have been subjected to high-pressure heating are softer than those that have not been treated, and have a texture similar to that of ready-made Nakashimanazuke.
Color tone: L*, a*, and b* were measured by the reflection method using a color difference meter (manufactured by Nippon Denshoku Co., Ltd.) for the color tone of the sample under each processing condition. As for the measurement results, as shown in FIG. 53, the green color of the leaves becomes considerably dull in proportion to the treatment temperature and treatment time. Further, FIG. 54(a) shows the color tone before the high pressure heating treatment, and (b) shows the color tone before the high pressure heating treatment.
Microbial hygiene test method: Suspend each sample in physiological saline, serially dilute, and add each diluted sample to standard agar medium (Eiken, for general viable cell count) and 0.01 (w/v) % chloramphenicol. Potato dextrose agar medium (Eiken, for fungi) was smeared (in the case of standard agar, the pour method was used). After culturing at 30 to 37° C. for 3 days, the number of microbial colonies formed was counted, and the number of viable general bacteria and fungi per 1 g of sample was calculated. With reference to the standards of the Food Sanitation Act, the standard values were 300 cfu/g or less for the general viable count and 1000 cfu/g or less for the fungal count. Non-detection was defined as 50 cfu/g or less for both general viable bacteria and fungi.
(Table 32A) shows the results of microbial hygiene inspection by treatment temperature (all treatment times are 30 minutes), and (Table 32B) shows the results of microbial hygiene inspection by treatment time (all treatment temperatures are 50°C).

In the table, “above the standard” means 300/g or more for general viable bacteria, 1000/g or more for fungi, and “below the standard” means 300/g or less for general viable bacteria, 1000/g or less for fungi, and not detected. refers to the limit of detection, 50/g or less.
As shown in the table, the number of viable bacteria is below the sanitary standard value for both treatment temperature and treatment time. This is thought to be due not only to the effect of high-pressure heating, as in radish, but also to the synergistic effect of the sterilization effect of the alcohol contained in the sake lees soaked in Nakashimana, which inhibited the growth of bacteria.
(3). Effect of treatment at 80 MPa or less and 100 MPa or more When the method for producing narazuke (pickled lees) of radish and Nakashimana was examined by changing only the pressure condition of the high pressure treatment condition in the range of 80 to 300 MPa, the results shown in Table 33 were obtained. It was confirmed that the invention is also effective in the pressure range of 80-300 MPa.

(3) Pickled radish in soy sauce / processing conditions Pressure: 1000 atmospheres (100 MPa)
Temperature: 40-70°C
Time: 15-60 minutes Production details Vegetables: Daikon radish (green neck)
Pickling liquid: 20% soy sauce aqueous solution (salinity concentration about 3.5%)
Production method: Peeled daikon radish was cut into a half-moon shape with a width of 1 cm. As a general method, a sample was prepared by removing the skin and cutting the daikon radish into half-moon shape with a thickness of 1 cm and immersing it in a 20% soy sauce aqueous solution at 5° C. for 24 hours.
- Evaluation method Salinity: each sample was pulverized and filtered, and the filtrate was used as a measurement sample and measured with a salinity meter (manufactured by Toa Denpa Co., Ltd.). As shown in FIG. 55, the measurement results showed that the salt concentration in the radish pulp was improved under all treatment conditions compared to the general method. In addition, as the treatment temperature rises, the ingredients permeate more and the salt concentration increases. Also, the salt concentration increases in proportion to the processing time.
Ease of biting off: each sample was prepared into a square cube of 1 cm high and 2 cm wide, and the stress when it was penetrated to 6 mm by a 5 mmφ cylindrical plunger was measured with a rheometer. As shown in FIG. 56, the measurement results show that at a treatment temperature of 60° C. or higher or for a treatment time of 60 minutes or longer, the texture becomes softer and easier to bite off. Raw texture can be maintained under processing conditions below them. However, softening was suppressed under any treatment conditions compared to the general method.
Color tone: L*, a*, and b* were measured by the reflection method using a color difference meter (manufactured by Nippon Denshoku Co., Ltd.) for the color tone of the sample under each processing condition. FIG. 57(a) shows the color tone before the high pressure heating treatment, and (b) shows the color tone before the high pressure heating treatment.
Degree of cell disruption: In this example, changes in the diffusion coefficient of water in vegetable tissue under high pressure treatment were examined. The diffusion coefficient of water was analyzed by NMR using radish treated under various conditions under high pressure as a sample. For NMR measurement, ESX400 (1H resonance frequency of 400 MHz) was used. The central portion of each sample was cut into a square of about 3 cm, rounded into a cylindrical shape, placed in an NMR measuring tube, and the diffusion coefficient of each sample was measured by changing the diffusion time from 0.1 to 1.0 sec by the PGSTE method. The results are shown in FIG. As shown in FIG. 58, the diffusion coefficient of water decreases significantly as the diffusion time increases when the range of movement within the tissue is small (limited diffusion). Looking at the figure, the range of decrease in the diffusion coefficient is smaller for all the high-pressure heating treated samples compared to the untreated sample, and the high-pressure heating treatment increases the range in which water can move in the tissue of the vegetable. In other words, it was confirmed that cell membrane destruction occurred. This makes it easier for the ingredients to penetrate.
Content of umami component: Each sample was suspended in physiological saline and thoroughly mixed. After removing the residue by centrifugation or filtering, amino acids in the sample were fluorescently labeled using AccQ-Fluor Reagent Kit (Waters). Each amino acid was eluted by high performance liquid chromatography (HPLC) with a linear gradient using the prescribed mobile phases (mobile phase A: AccQ-Eluent A (Waters), mobile phase B: 60 (v/v) % acetonitrile). . AccQ-tag Amino Acid Analysis Column (Waters) was used as the column, and the column oven was set at 40°C and elution was performed at a flow rate of 1 ml/min. Upon detection, fluorescence intensity was measured at an excitation wavelength of 250 nm and a fluorescence wavelength of 395 nm. As shown in FIG. 59, when the high-pressure heating treatment is performed, the umami component increases in proportion to the increase in treatment temperature. It is thought that this is because the umami component of soy sauce permeated the flesh of the radish.
Microbial hygiene test method: Suspend each sample in physiological saline, serially dilute, and add each diluted sample to standard agar medium (Eiken, for general viable cell count) and 0.01 (w/v) % chloramphenicol. Potato dextrose agar medium (Eiken, for fungi) was smeared (in the case of standard agar, the pour method was used). After culturing at 30 to 37° C. for 3 days, the number of microbial colonies formed was counted, and the number of viable general bacteria and fungi per 1 g of the sample was calculated. With reference to the standards of the Food Sanitation Act, the standard values were 300 cfu/g or less for general viable counts and 1000 cfu/g or less for fungal counts. Non-detection was defined as 50 cfu/g or less for both general viable bacteria and fungi.
(Table 34A) shows the results of microbial hygiene inspections by treatment temperature (all treatment times were 3 hours), and (Table 34B) shows the results of microbial hygiene inspections by treatment time (all treatment temperatures were 70°C).

In the table, above the standard means 100,000/g or more for general viable bacteria, 1000/g or more for fungi, and below the standard means 100,000/g or less for general viable bacteria, 1000/g or less for fungi, and no Detection refers to the limit of detection, 50/g or less.
As shown in the table, when high-pressure heating treatment is performed at each temperature for 30 minutes, the sanitation standard value is exceeded at 40°C after one month, but at 50°C, general viable bacteria are below the sanitation standard value, and fungi It was not detected. In addition, it can be completely sterilized at a treatment temperature of 60°C or higher. There was no change in the number of bacteria with time after high-pressure heating treatment at 50°C, but all were below the sanitary standard value.

(4) Nakashima greens pickled in soy sauce / processing conditions Pressure: 1000 atmospheres (100 MPa)
Temperature: 40-70°C
Time: 5 to 30 minutes Production details Vegetables: Nakashima greens Pickling liquid: 40% soy sauce aqueous solution (salinity concentration about 7.3%)
Production method: The ratio of stems and leaves of Nakashima greens was set to 1:1, and the whole weight and the same amount of pickling liquid were placed in a bag and vacuum-packaged.
- Evaluation method Salinity: each sample was pulverized and filtered, and the filtrate was used as a measurement sample and measured with a salinity meter (manufactured by Toa Denpa Co., Ltd.). As shown in FIG. 60, the measurement results show that the ingredients permeate more and the salt concentration increases in proportion to the increase in treatment temperature. Also, the salt concentration increases in proportion to the processing time.
Hardness: Each sample is adjusted to a stem cut to 1.5 cm to 2 cm in length, the stem is cut with a needle-like plunger of 2 mmφ, and the stress when it penetrates to 10 mm so as to penetrate the center is measured by a rheometer. It was measured. As shown in FIG. 61, the measurement results show that the hardness of the stalks of soy sauce-pickled Nakashima greens that have been subjected to high-pressure heating is softer than that of the untreated ones, but the texture is more chewy than that of ready-made Nakashima greens. Become.
Color tone: L*, a*, and b* were measured by the reflection method using a color difference meter (manufactured by Nippon Denshoku Co., Ltd.) for the color tone of the sample under each processing condition. As for the measurement results, as shown in FIG. 62, the green color of the leaves becomes dull in proportion to the treatment temperature and treatment time. This is proportional to the amount of soy sauce (salt content) soaked in. Also, FIG. 63(a) shows the color tone before the high pressure heating treatment, and (b) shows the color tone before the high pressure heating treatment.
Microbial hygiene test method: Suspend each sample in physiological saline, serially dilute, and add each diluted sample to standard agar medium (Eiken, for general viable cell count) and 0.01 (w/v) % chloramphenicol. Potato dextrose agar medium (Eiken, for fungi) was smeared (in the case of standard agar, the pour method was used). After culturing at 30 to 37° C. for 3 days, the number of microbial colonies formed was counted, and the number of viable general bacteria and fungi per 1 g of sample was calculated. With reference to the standards of the Food Sanitation Act, the standard values were 300 cfu/g or less for the general viable count and 1000 cfu/g or less for the fungal count. Non-detection was defined as 50 cfu/g or less for both general viable bacteria and fungi.
(Table 35A) shows the results of microbial hygiene inspection by treatment temperature (all treatment times are 15 minutes), and (Table 35B) shows the results of microbial hygiene inspection by treatment time (all treatment temperatures are 60°C).

In the table, above the standard means 100,000/g or more for general viable bacteria, 1000/g or more for fungi, and below the standard means 100,000/g or less for general viable bacteria, 1000/g or less for fungi, and no Detection refers to the limit of detection, 50/g or less.
As shown in the table, general viable bacteria were detected at all treatment temperatures, but at treatment temperatures of 50°C or higher in high-pressure heating treatment, they were below the sanitary standard value. For fungi, the treatment temperature of high-pressure heating treatment is 50°C or higher, exceeding the sanitary standard value. When high-pressure heating treatment was performed at 60°C, no change in the number of viable bacteria was observed depending on the treatment time.

(5) Effect of treatment at 80 MPa or less and 100 MPa or more When the method for pickling radish and Nakashima greens in soy sauce was examined by changing only the pressure condition of the high pressure treatment condition in the range of 80 to 300 MPa, the results are shown in Table 36. It was confirmed that the present invention is effective even in the pressure range of 80-300 MPa.

Additional experiment (fruit)
(1) Plum pickled in syrup/treatment conditions Pressure: 1000 atmospheres (100 MPa)
Temperature: 40-70°C
Time: 15 to 60 minutes Production details Fruit: Frozen plums from Ishikawa Prefecture (Ishikawa No. 1)
Syrup: 60% granulated sugar water (50.4% sugar content)
Production method: 5 plums were placed in a bag together with twice the amount of syrup and vacuum-packaged.
・Evaluation method Sugar content: Each sample was pulverized and filtered, and the filtrate was used as a measurement sample and measured with a sugar content meter (manufactured by Atago Co., Ltd.). As shown in FIG. 64, the measurement results confirmed that the higher the treatment temperature and the longer the treatment time at 60°C, the higher the sugar content of the syrup and the easier it penetrates into the pulp. .
Acidity: The high-pressure heat-treated sample was taken out of the bag, the syrup was wiped off, the seeds were removed, and the sample was homogenized with a mixer. A sample that was made into a paste by homogenization was used as a sample. The acidity (%) of the sample was measured using an acidity titrator, and was converted into the equivalent amount of citric acid for evaluation. As shown in FIG. 65, the measurement results show that the higher the treatment temperature and the longer the treatment time at 60° C., the lower the acidity of the pulp and the easier the acid elution from the pulp. confirmed.
Hardness: One piece of frozen plum was fixed on a measuring table, and the stress when it was compressed to 6 mm with a 3 mmφ cylindrical plunger was measured with a rheometer. As shown in FIG. 66, the measurement results show that when the temperature is 60° C. or higher and the high-pressure heating treatment is performed at that temperature for 30 minutes or longer, the texture of the pulp becomes soft.
Color tone: L*, a*, and b* were measured by the reflection method using a color difference meter (manufactured by Nippon Denshoku Co., Ltd.) for the color tone of the sample under each processing condition. As shown in FIG. 67, the measurement results show that the color of the pericarp becomes slightly dull due to the high-pressure heating treatment, but there is no problem with marketability. Also, FIG. 68(a) shows the color tone before high pressure heating treatment, and FIG. 68(b) shows the color tone before high pressure heating treatment.
Microbial hygiene test method: Suspend each sample in physiological saline, serially dilute, and add each diluted sample to standard agar medium (Eiken, for general viable cell count) and 0.01 (w/v) % chloramphenicol. Potato dextrose agar medium (Eiken, for fungi) was smeared (in the case of standard agar, the pour method was used). After culturing at 30 to 37° C. for 3 days, the number of microbial colonies formed was counted, and the number of viable general bacteria and fungi per 1 g of sample was calculated. With reference to the standards of the Food Sanitation Act, the standard values were 300 cfu/g or less for the general viable count and 1000 cfu/g or less for the fungal count. Non-detection was defined as 50 cfu/g or less for both general viable bacteria and fungi.
(Table 37A) shows the results of microbial hygiene inspection by treatment temperature (all treatment times are 30 minutes), and (Table 37B) shows the results of microbial hygiene inspection by treatment time (all treatment temperatures are 60°C).

In the table, “above the standard” means 300/g or more for general viable bacteria, 1000/g or more for fungi, and “below the standard” means 300/g or less for general viable bacteria, 1000/g or less for fungi, and not detected. refers to the limit of detection, 50/g or less.
As shown in the table, it was confirmed that both general viable bacteria and fungi could be sterilized by high-pressure heating at 60°C or higher, with no detection in most of the test plots. Moreover, when the treatment temperature of the high-pressure heating treatment is 60° C., the treatment time is 30 minutes or longer, and long-term storage is possible.

(2) Japanese pear soaked in syrup/treatment conditions Pressure: 1000 atmospheres (100 MPa)
Temperature: 40-70°C
Time: 15-60 minutes Production details Fruit: Japanese pear (Yosui)
Syrup: 15% granulated sugar water (14.3% sugar content)
Method: The peeled and seeded pears were bagged with an equal weight of syrup and vacuum packed.
・Evaluation method Sugar content: Each sample was pulverized and filtered, and the filtrate was used as a measurement sample and measured with a sugar content meter (manufactured by Atago Co., Ltd.). As shown in FIG. 69, the measurement results show that when the high-pressure heating treatment is performed, the components of the syrup permeate into the pulp in proportion to the treatment temperature and treatment time.
Hardness: Each sample was prepared into a square cube with a height of 1 cm and a width of 2 cm, and the stress was measured by a rheometer when a cylindrical plunger with a diameter of 5 mm penetrated to a depth of 6 mm. As shown in FIG. 70, the measurement results show that when the temperature is 60° C. or higher and the treatment time is long, the texture of the pulp becomes slightly softer, but the texture of the untreated pulp can be almost maintained.
Color tone: L*, a*, and b* were measured by the reflection method using a color difference meter (manufactured by Nippon Denshoku Co., Ltd.) for the color tone of the sample under each processing condition. As shown in FIG. 71, the measurement results show translucency due to the permeation of the syrup, but there is almost no change in color due to treatment conditions. Moreover, this color tone does not pose any problem in marketability. Also, FIG. 72(a) shows the color tone before the high pressure heating treatment, and (b) shows the color tone before the high pressure heating treatment.
Degree of cell disruption: In this example, changes in the diffusion coefficient of water in vegetable tissue under high pressure treatment were examined. The diffusion coefficient of water was analyzed by NMR using pears treated under various conditions under high pressure as samples. For NMR measurement, ESX400 (1H resonance frequency of 400 MHz) was used. The central portion of each sample was cut into a square of about 3 cm, rounded into a cylindrical shape, placed in an NMR measuring tube, and the diffusion coefficient of each sample was measured by changing the diffusion time from 0.1 to 1.0 sec by the PGSTE method. As shown in FIG. 73, the high-pressure heating process destroys the cells in proportion to the treatment temperature.

(3) Grapes soaked in syrup/treatment conditions Pressure: 1000 atmospheres (100 MPa)
Temperature: 40-70°C
Time: 15-60 minutes Production details Fruit: Ishikawa Prefecture grapes (Ruby Roman)
Syrup: 20% granulated sugar water (18.9% sugar content)
Method: 5 peeled grapes were bagged with their 40% amount of syrup and vacuum packed.
・Evaluation method Sugar content: Each sample was pulverized and filtered, and the filtrate was used as a measurement sample and measured with a sugar content meter (manufactured by Atago Co., Ltd.). As shown in FIG. 74, the measurement results show that when grapes are subjected to high-pressure heating treatment, the sugar content of the syrup permeates into the pulp to the same extent regardless of the treatment temperature and treatment time.
Hardness and ease of biting: A single grape was fixed on a measuring table and the stress when compressed to 6 mm with a 3 mmφ cylindrical plunger was measured with a rheometer. As shown in FIG. 75, the measurement results show that the hardness of the grapes and the ease of chewing through the grapes are the same as those of the untreated grapes even after the high-pressure heating treatment, and the raw texture can be maintained.
Color tone: L*, a*, and b* were measured by the reflection method using a color difference meter (manufactured by Nippon Denshoku Co., Ltd.) for the color tone of the sample under each processing condition. As shown in FIG. 76, the measurement results show that when the color difference is compared with the color of the untreated pulp, the color of the grape pulp does not change even after the high-pressure heating treatment, and the color of the raw grape pulp is maintained. be able to. FIG. 77(a) shows the color tone before the high pressure heating treatment, and (b) shows the color tone before the high pressure heating treatment.
Microbial hygiene test method: Suspend each sample in physiological saline, serially dilute, and add each diluted sample to standard agar medium (Eiken, for general viable cell count) and 0.01 (w/v) % chloramphenicol. Potato dextrose agar medium (Eiken, for fungi) was smeared (in the case of standard agar, the pour method was used). After culturing at 30 to 37° C. for 3 days, the number of microbial colonies formed was counted, and the number of viable general bacteria and fungi per 1 g of sample was calculated. With reference to the standards of the Food Sanitation Act, the standard values were 300 cfu/g or less for the general viable count and 1000 cfu/g or less for the fungal count. Non-detection was defined as 50 cfu/g or less for both general viable bacteria and fungi.
(Table 38A) shows the results of microbial hygiene inspection by treatment temperature (all treatment times are 30 minutes), and (Table 38B) shows the results of microbial hygiene inspection by treatment time (all treatment temperatures are 60°C).

In the table, “above the standard” means 300/g or more for general viable bacteria, 1000/g or more for fungi, and “below the standard” means 300/g or less for general viable bacteria, 1000/g or less for fungi, and not detected. refers to the limit of detection, 50/g or less.
As shown in the table, by treatment temperature of high-pressure heat treatment, both general viable bacteria and fungi were below the standard or not detected at any temperature. At treatment temperatures of 50° C. and above, no common viable bacteria and fungi were detected until after 6 months. Moreover, when the treatment temperature of the high-pressure heat treatment was 60° C., the treatment time was 30 minutes or longer, and no viable bacteria or fungi were detected.

(4). Effect of treatment at 80 MPa or less and 100 MPa or more When the syrup pickling method for ume, Japanese pear, and grapes was examined by changing only the pressure condition of the high-pressure treatment condition in the range of 80 to 300 MPa, the results shown in Table 39 were obtained. is effective even in the pressure range of 80 to 300 MPa.

Additional experiment (beef)
(1) Beef (outer thigh) marinated in miso and processing conditions Pressure: 1000 atmospheres (100 MPa)
Temperature: 20°C to 60°C
Time: 7.5 hours to 30 hours Production content Meat: Beef (outer thigh)
Pickles: seasoned miso (78% miso, 11% sugar, 11% mirin)
Production method: A 1 cm-thick slice of beef thigh was coated with 1/2 of pickled bed, placed in a bag, and vacuum-packaged. As a general production method, beef thighs were wrapped in gauze, coated with 1/2 of the amount of pickles (seasoned miso), placed in a bag, vacuum-packaged, and stored at 5°C for 3 days.
・Evaluation method Sugar composition: A sample subjected to high-pressure heating was taken out from the bag, and after wiping off the seasoning liquid, etc., it was homogenized with a mixer. After adding 1 ml of an 8% trichloroacetic acid solution to 1 g of the homogenized sample and stirring vigorously, the extract was filtered and diluted to 10 ml with distilled water to obtain a sample. An HPLC system (Shimadzu Corporation) was used for analysis. The column was Mightysil NH ₂ (5 μm) (Kanto Kagaku Co., Ltd.), the detector was a differential refractometer (RID-10A), the mobile phase was 70% acetonitrile, and the analysis was performed at a flow rate of 1 ml/min. Three components, fructose, glucose, and sucrose, were evaluated by measuring the sugar composition of samples and quantifying them. As shown in FIG. 78, the sugar concentration permeated the sample treated with high pressure heating for 15 hours more than 10 times higher than that of the untreated sample regardless of the treatment temperature. In addition, the permeation of the sugar components contained in the marinade into the beef thigh increases in proportion to the rise in the treatment temperature, and at a treatment temperature of 60°C, the concentration is higher than the general method.
Hardness: Each sample was prepared into a cube of 1 cm square, and a razor-blade plunger was applied perpendicularly to muscle fibers, and the stress was measured by a rheometer when the muscle fibers were penetrated to a depth of 8 mm. As for the measurement results, as shown in FIG. 79, the hardness after heating becomes softer by performing the high-pressure heating treatment as compared with the untreated and the general method. It is thought that the treatment at 60°C accelerated the enzymatic reaction and made it softer. In the treatment at 40°C, the hardness did not change much with the treatment time, so it is considered that the treatment time of 7.5 hours is sufficient.
Color tone: Fig. 80(a) shows the color tone before the high pressure heating treatment, and (b) shows the color tone before the high pressure heating treatment.
Content of umami components: Suspend each sample in physiological saline, mix well, and eluate each amino acid by high performance liquid chromatography (HPLC) using either centrifugation or a linear gradient using a filter (60 (v/v) % acetonitrile). bottom. AccQ-tag Amino Acid Analysis as a column
s Column (Waters) was used, the column oven was set at 40°C, and elution was performed at a flow rate of 1 ml/min. Upon detection, fluorescence intensity was measured at an excitation wavelength of 250 nm and a fluorescence wavelength of 395 nm. As shown in FIG. 81, the measurement results show that when the high-pressure heating treatment is performed, the amount of umami component increases in proportion to the treatment temperature and treatment time, especially at a treatment temperature of 60°C.
Microbial hygiene test method: Suspend each sample in physiological saline, serially dilute, and add each diluted sample to standard agar medium (Eiken, for general viable cell count) and 0.01 (w/v) % chloramphenicol. Potato dextrose agar medium (Eiken, for fungi) was smeared (in the case of standard agar, the pour method was used). After culturing at 30 to 37° C. for 3 days, the number of microbial colonies formed was counted, and the number of viable general bacteria and fungi per 1 g of sample was calculated. With reference to the standards of the Food Sanitation Act, the standard values were 300 cfu/g or less for the general viable count and 1000 cfu/g or less for the fungal count. Non-detection was defined as 50 cfu/g or less for both general viable bacteria and fungi.
(Table 40A) shows the results of the microbial hygiene inspection by treatment temperature (all treatment times are 15 hours), and (Table 40B) shows the results of the microbial hygiene inspection by treatment time (all treatment temperatures are 40°C).

In the table, above the standard means 100,000/g or more for general viable bacteria, 1000/g or more for fungi, and below the standard means 100,000/g or less for general viable bacteria, 1000/g or less for fungi, and no Detection refers to the limit of detection, 50/g or less.
As shown in the table, general viable bacteria were detected for each treatment temperature and treatment time, but all were below the sanitary standard values. No fungi were detected at any treatment temperature or treatment time. Beef thigh marinated in miso can be stored at 5°C for one month under any processing conditions.

(2) Beef (tongue) marinated in miso and processing conditions Pressure: 1000 atmospheres (100 MPa)
Temperature: 20°C to 60°C
Time: 7.5 hours to 30 hours Production content Meat: Beef tongue (crown cut)
Pickles: seasoned miso (78% miso, 11% sugar, 11% mirin)
Production method: A 1 cm-thick slice of beef thigh was coated with 1/2 of pickled bed, placed in a bag, and vacuum-packaged. As a general production method, the beef tongue was wrapped in gauze, coated with 1/2 of the amount of pickles (seasoned miso) of the ingredients, placed in a bag, vacuum-packaged, and stored at 5°C for 3 days.
・Evaluation method Sugar composition: A sample subjected to high-pressure heating was taken out from the bag, and after wiping off the seasoning liquid, etc., it was homogenized with a mixer. After adding 1 ml of an 8% trichloroacetic acid solution to 1 g of the homogenized sample and stirring vigorously, the extract was filtered and diluted to 10 ml with distilled water to obtain a sample. An HPLC system (Shimadzu Corporation) was used for analysis. The column was Mightysil NH ₂ (5 μm) (Kanto Kagaku Co., Ltd.), the detector was a differential refractometer (RID-10A), the mobile phase was 70% acetonitrile, and the analysis was performed at a flow rate of 1 ml/min. Three components, fructose, glucose, and sucrose, were evaluated by measuring the sugar composition of samples and quantifying them. As shown in FIG. 82, the measurement results show that the sugar concentration of about 3 g or more permeated the sample treated with high pressure heating for 15 hours compared to the untreated sample, regardless of the treatment temperature. In addition, the permeation of the sugar components contained in the marinade into the beef tongue increases in proportion to the rise in the treatment temperature, and at a treatment temperature of 40°C or higher, the concentration is higher than the general method.
Hardness: Each sample was prepared into a cube of 1 cm square, and a razor-blade plunger was applied perpendicularly to muscle fibers, and the stress was measured by a rheometer when the muscle fibers were penetrated to a depth of 8 mm. As for the measurement results, as shown in FIG. 83, the hardness after heating becomes softer by performing the high-pressure heating treatment as compared with the untreated and the general method. It is thought that the treatment at 60°C accelerated the enzymatic reaction and made it softer. In the 40°C treatment, the longer the treatment time, the softer it becomes.
Color tone: Fig. 84(a) shows the color tone before the high pressure heating treatment, and (b) shows the color tone before the high pressure heating treatment.
Content of umami component: Each sample was suspended in physiological saline and thoroughly mixed. After removing the residue by centrifugation or filtering, amino acids in the sample were fluorescently labeled using AccQ-Fluor Reagent Kit (Waters). Each amino acid was eluted by high performance liquid chromatography (HPLC) with a linear gradient using the prescribed mobile phases (mobile phase A: AccQ-Eluent A (Waters), mobile phase B: 60 (v/v) % acetonitrile). . AccQ-tag Amino Acid Analysis Column (Waters) was used as the column, and the column oven was set at 40°C and elution was performed at a flow rate of 1 ml/min. Upon detection, fluorescence intensity was measured at an excitation wavelength of 250 nm and a fluorescence wavelength of 395 nm. As shown in FIG. 85, the measurement results show that the amount of umami component is greatly increased at a treatment temperature of 60°C. Moreover, the amount of umami components increases in proportion to the processing time.
Microbial hygiene test method: Suspend each sample in physiological saline, serially dilute, and add each diluted sample to standard agar medium (Eiken, for general viable cell count) and 0.01 (w/v) % chloramphenicol. Potato dextrose agar medium (Eiken, for fungi) was smeared (in the case of standard agar, the pour method was used). After culturing at 30 to 37° C. for 3 days, the number of microbial colonies formed was counted, and the number of viable general bacteria and fungi per 1 g of sample was calculated. With reference to the standards of the Food Sanitation Act, the standard values were 300 cfu/g or less for the general viable count and 1000 cfu/g or less for the fungal count. Non-detection was defined as 50 cfu/g or less for both general viable bacteria and fungi.
(Table 41A) shows the results of the microbial hygiene inspection by treatment temperature (all treatment times are 15 hours), and (Table 41B) shows the results of the microbial hygiene inspection by treatment time (all treatment temperatures are 40°C).

In the table, above the standard means 100,000/g or more for general viable bacteria, 1000/g or more for fungi, and below the standard means 100,000/g or less for general viable bacteria, 1000/g or less for fungi, and no Detection refers to the limit of detection, 50/g or less.
As shown in the table, general viable bacteria were detected for each treatment temperature and treatment time, but all were below the sanitary standard values. No fungi were detected at any treatment temperature or treatment time. Miso-pickled beef tongue can be stored at 5°C for one month under any treatment conditions, as with beef thigh.

(3) Beef (outer thigh) pickled in rice malt and processing conditions Pressure: 1000 atmospheres (100 MPa)
Temperature: 20°C to 60°C
Time: 1 to 5 hours Production content Meat: Beef (outer thigh)
Pickling liquid: rice koji water (koji: water = 1:1)
Production method: Beef thigh meat sliced into 1 cm thickness and 1/5 amount of rice koji water were placed in a bag and vacuum-packaged.
Evaluation method glucose concentration: After homogenizing the meat after each treatment, 5 ml of 8% trichloroacetic acid aqueous solution was added to 5 g of the sample, vigorously shaken, filtered, and clarified. The glucose concentration of the constant-volume liquid was measured using a glucose CII test manufactured by Wako Pure Chemical Industries, Ltd., and converted to the glucose concentration contained in the original sample. Three replicates were performed per treatment, and the mean and standard deviation were determined. As for the measurement results, as shown in FIG. 86, the glucose concentration in the beef thigh increased to 200 mg or more, regardless of the treatment time and temperature of the high-pressure heating treatment, compared to the untreated beef. In addition, the glucose concentration of the beef thigh increases in proportion to the increase in treatment temperature. Also, the glucose concentration increases in proportion to the length of treatment time. In particular, since the processing temperature of 60°C is the optimal temperature for koji enzymes, the increase in glucose concentration is remarkable.
Hardness: Each sample was prepared into a cube of 1 cm square, and a razor-blade plunger was applied perpendicularly to muscle fibers, and the stress was measured by a rheometer when the muscle fibers were penetrated to a depth of 8 mm. As shown in FIG. 87, the measurement results show that the hardness of the treated meat was softened compared to the untreated meat regardless of the treatment time and temperature of the high pressure heating treatment. In addition, there was a tendency of softening as the treatment temperature increased. In particular, at a treatment temperature of 60°C, which is the optimum temperature for koji enzymes, the hardness after heating becomes softer. The hardness after heating becomes softer in proportion to the length of treatment time at 60°C.
Color tone: Fig. 88(a) shows the color tone before the high pressure heating treatment, and (b) shows the color tone before the high pressure heating treatment.
Content of umami component: Each sample was suspended in physiological saline and thoroughly mixed. After removing the residue by centrifugation or filtering, amino acids in the sample were fluorescently labeled using AccQ-Fluor Reagent Kit (Waters). Each amino acid was eluted by high performance liquid chromatography (HPLC) with a linear gradient using the prescribed mobile phases (mobile phase A: AccQ-Eluent A (Waters), mobile phase B: 60 (v/v) % acetonitrile). . AccQ-tag Amino Acid Analysis Column (Waters) was used as the column, and the column oven was set at 40°C and elution was performed at a flow rate of 1 ml/min. Upon detection, fluorescence intensity was measured at an excitation wavelength of 250 nm and a fluorescence wavelength of 395 nm. As shown in FIG. 89, the measurement results show that when the high-pressure heating treatment is performed, the amount of umami component increases in proportion to the treatment temperature and treatment time. In addition, when the treatment time was 15 hours, the treatment temperature was 40°C or higher, and when the treatment temperature was 60°C, the umami component content was 1% or higher when the treatment time was 15 hours or longer. Test method: Suspend each sample in physiological saline and serially dilute each diluted sample on a standard agar medium (Eiken, for general viable cell count) and potato containing 0.01 (w/v) % chloramphenicol. It was smeared on a dextrose agar medium (Eiken, for fungi) (in the case of standard agar, the pour method was used). After culturing at 30-37°C for 3 days, the number of microbial colonies formed was counted, and the number of viable general bacteria and fungi per 1 g of sample was calculated. With reference to the standards of the Food Sanitation Act, the standard values were 300 cfu/g or less for the general viable count and 1000 cfu/g or less for the fungal count. Non-detection was defined as 50 cfu/g or less for both general viable bacteria and fungi.
(Table 42A) shows the results of the microbial hygiene inspection by treatment temperature (all treatment times were 15 hours), and (Table 42B) shows the results of the microbial hygiene inspection by treatment time (all treatment temperatures were 40°C).

In the table, above the standard means 100,000/g or more for general viable bacteria, 1000/g or more for fungi, and below the standard means 100,000/g or less for general viable bacteria, 1000/g or less for fungi, and no Detection refers to the limit of detection, 50/g or less.
As shown in the table, by treatment temperature, both general viable bacteria and fungi exceeded the sanitary standard values when treated at 40°C or lower, but were not detected at 60°C or higher. By treatment time at 60°C, viable bacteria were detected within a range that did not exceed the sanitation standard value after one month only in those treated for 1 hour, but there was almost no difference in the number of viable bacteria depending on the treatment time. Considering that the active temperature of koji enzymes is 60°C, we recommend high-pressure heating at 60°C or higher for pickling beef thighs in koji.

(4) Beef (tongue) marinated in rice malt and processing conditions Pressure: 1000 atmospheres (100 MPa)
Temperature: 20°C to 60°C
Time: 1 to 5 hours Production details Meat: Beef tongue (before tongue)
Pickling liquid: rice koji water (koji: water = 1:1)
Production method: Beef thigh meat sliced into 1 cm thickness and 2/5 amount of rice koji water were placed in a bag and vacuum-packaged.
Evaluation method glucose concentration: After homogenizing the meat after each treatment, 5 ml of 8% trichloroacetic acid aqueous solution was added to 5 g of the sample, vigorously shaken, filtered, and clarified. The glucose concentration of the constant-volume liquid was measured using a glucose CII test manufactured by Wako Pure Chemical Industries, Ltd., and converted to the glucose concentration contained in the original sample. Three replicates were performed per treatment, and the mean and standard deviation were determined. As for the measurement results, as shown in FIG. 90, the glucose concentration in the beef tongue improved to about 200 mg or more, regardless of the treatment time and temperature of the high-pressure heating treatment, compared to the untreated beef tongue. In addition, the glucose concentration in beef tongue increases in proportion to the increase in treatment temperature. Also, the glucose concentration increases in proportion to the length of treatment time. In particular, since the processing temperature of 60°C is the optimal temperature for koji enzymes, the increase in glucose concentration is remarkable.
Hardness: Each sample was prepared into a cube of 1 cm square, and a razor-blade plunger was applied perpendicularly to muscle fibers, and the stress was measured by a rheometer when the muscle fibers were penetrated to a depth of 8 mm. As shown in FIG. 91, the measurement results show that the hardness of the treated meat was softened compared to the untreated meat regardless of the treatment time and temperature of the high-pressure heating treatment. In addition, the softening was proportional to the increase in treatment temperature and the length of treatment time.
Color tone: Fig. 92(a) shows the color tone before the high pressure heating treatment, and (b) shows the color tone before the high pressure heating treatment.
Content of umami component: Each sample was suspended in physiological saline and thoroughly mixed. After removing the residue by centrifugation or filtering, amino acids in the sample were fluorescently labeled using AccQ-Fluor Reagent Kit (Waters). Each amino acid was eluted by high performance liquid chromatography (HPLC) with a linear gradient using the prescribed mobile phases (mobile phase A: AccQ-Eluent A (Waters), mobile phase B: 60 (v/v) % acetonitrile). . AccQ-tag Amino Acid Analysis Column (Waters) was used as the column, and the column oven was set at 40°C and elution was performed at a flow rate of 1 ml/min. Upon detection, fluorescence intensity was measured at an excitation wavelength of 250 nm and a fluorescence wavelength of 395 nm. As shown in FIG. 93, the measurement results show that when the high-pressure heating treatment is performed, the amount of umami component increases in proportion to the treatment temperature and treatment time. Also, when the treatment time was 15 hours, the treatment temperature was 20°C or higher, and when the treatment temperature was 60°C, the umami component content was 1% or more when the treatment time was 15 hours or longer.
Microbial hygiene test method: Suspend each sample in physiological saline, serially dilute, and add each diluted sample to standard agar medium (Eiken, for general viable cell count) and 0.01 (w/v) % chloramphenicol. Potato dextrose agar medium (Eiken, for fungi) was smeared (in the case of standard agar, the pour method was used). After culturing at 30 to 37° C. for 3 days, the number of microbial colonies formed was counted, and the number of viable general bacteria and fungi per 1 g of sample was calculated. With reference to the standards of the Food Sanitation Act, the standard values were 300 cfu/g or less for the general viable count and 1000 cfu/g or less for the fungal count. Non-detection was defined as 50 cfu/g or less for both general viable bacteria and fungi.
(Table 43A) shows the results of microbial hygiene inspection by treatment temperature (all treatment times are 3 hours), and (Table 43B) shows the results of microbial hygiene inspection by treatment time (all treatment temperatures are 60°C).

In the table, above the standard means 100,000/g or more for general viable bacteria, 1000/g or more for fungi, and below the standard means 100,000/g or less for general viable bacteria, 1000/g or less for fungi, and no Detection refers to the limit of detection, 50/g or less.
As shown in the table, by treatment temperature, both general viable bacteria and fungi exceeded the sanitary standard values when treated at 40°C or lower, but fell below the sanitary standard values when treated at 60°C or higher. By treatment time at 60°C, general viable bacteria were detected within a range that did not exceed the sanitary standard value at any treatment time, but there was no difference in the number of viable bacteria depending on the treatment time. As with beef thigh meat, the enzyme activity temperature of koji is 60°C, so we recommend high-pressure heating at 60°C or higher for pickling beef tongue in koji.

(5) Effect of treatment at 80 MPa or less and 100 MPa or more When the method for producing beef miso and koji pickles was examined by changing only the pressure condition of the high pressure treatment condition in the range of 80 to 300 MPa, the results shown in Table 44 were obtained. It was confirmed that the invention is also effective in the pressure range of 80-300 MPa.

Additional experiment (hard tofu)
(1) Hard tofu pickled in lees and processing conditions Pressure: 1000 atmospheres (100 MPa)
Temperature: 20°C to 60°C
Time: 7.5 hours to 30 hours Production details Processed product: Hard tofu Pickled bed: Seasoning lees (40% sake lees, 40% miso, 10% sake, 10% mirin)
Production method: Hard tofu cut into 1 cm wide pieces was coated with 1/2 amount of seasoned sake lees, placed in a bag, and vacuum-packaged.
Evaluation method glucose concentration: After homogenizing the fish meat after each treatment, 5 ml of 8% trichloroacetic acid aqueous solution was added to 5 g of the sample, vigorously shaken, filtered and clarified. The glucose concentration of the constant-volume liquid was measured using a glucose CII test manufactured by Wako Pure Chemical Industries, Ltd., and converted to the glucose concentration contained in the original sample. Three replicates were performed per treatment, and the mean and standard deviation were determined. As shown in FIG. 94, the measurement results show that the high-pressure heating process increases the glucose concentration. It is particularly high at the treatment temperature of 40°C, and the glucose concentration increases in proportion to the treatment time.
Hardness: Each sample was prepared into a 1 cm square cube, and the stress when it was compressed by 80% with a 10 mmφ cylindrical plunger was measured with a rheometer. As shown in FIG. 95, the hardness of hard tofu is softened by high-pressure heating. Like the glucose concentration, it becomes particularly soft at a treatment temperature of 40°C, and becomes softer in proportion to the treatment time.
Color tone: L*, a*, and b* were measured by the reflection method using a color difference meter (manufactured by Nippon Denshoku Co., Ltd.) for the color tone of the sample under each processing condition. As shown in FIG. 96, the measurement results show that the color changes in proportion to the treatment temperature and treatment time, becoming more yellow. FIG. 97(a) shows the color tone before the high pressure heating treatment, and (b) shows the color tone before the high pressure heating treatment.
Content of umami component: Each sample was suspended in physiological saline and thoroughly mixed. After removing the residue by centrifugation or filtering, amino acids in the sample were fluorescently labeled using AccQ-Fluor Reagent Kit (Waters). Each amino acid was eluted by high performance liquid chromatography (HPLC) with a linear gradient using the prescribed mobile phases (mobile phase A: AccQ-Eluent A (Waters), mobile phase B: 60 (v/v) % acetonitrile). . AccQ-tag Amino Acid Analysis Column (Waters) was used as the column, and the column oven was set at 40°C and elution was performed at a flow rate of 1 ml/min. Upon detection, fluorescence intensity was measured at an excitation wavelength of 250 nm and a fluorescence wavelength of 395 nm. As shown in FIG. 98, the measurement results show that the high-pressure heating treatment increases the amount of umami components, especially at a treatment temperature of 60°C.
Microbial hygiene test method: Suspend each sample in physiological saline, serially dilute, and add each diluted sample to standard agar medium (Eiken, for general viable cell count) and 0.01 (w/v) % chloramphenicol. Potato dextrose agar medium (Eiken, for fungi) was smeared (in the case of standard agar, the pour method was used). After culturing at 30-37°C for 3 days, the number of microbial colonies formed was counted, and the number of viable general bacteria and fungi per 1 g of sample was calculated. With reference to the standards of the Food Sanitation Act, the standard values were 300 cfu/g or less for the general viable count and 1000 cfu/g or less for the fungal count. Non-detection was defined as 50 cfu/g or less for both general viable bacteria and fungi.
(Table 45A) shows the results of microbial hygiene inspection by treatment temperature (all treatment times are 15 hours), and (Table 45B) shows the results of microbial hygiene inspections by treatment time (all treatment temperatures are 40°C).

In the table, above the standard means 100,000/g or more for general viable bacteria, 1000/g or more for fungi, and below the standard means 100,000/g or less for general viable bacteria, 1000/g or less for fungi, and no Detection refers to the limit of detection, 50/g or less.
As shown in the table, the number of viable bacteria is below the sanitary standard value for both treatment temperature and treatment time. This is thought to be due to the synergistic effect of not only the effect of high-pressure heating, but also the sterilization effect of the alcohol contained in the sake lees soaked into the hard tofu, which inhibits the growth of bacteria.

(2) Effect of treatment at 80 MPa or less and 100 MPa or more When the method for producing hard tofu pickles was examined by changing only the pressure condition of the high pressure treatment condition in the range of 80 to 300 MPa, the results shown in Table 46 were obtained. It was confirmed that it is effective even in the pressure range of 80 to 300 MPa.

Additional experiment (seafood)
(1) Fukurugi lees pickling/treatment conditions Pressure: 1000 atmospheres (100 MPa)
Temperature: 20°C to 60°C
Time: 7.5 hours to 30 hours Production details Seafood: Fukuragi pickled bed: Seasoning lees (73% sake lees, 11.2% powdered rice koji, 7.5% sake, 7.5% mirin, 0.8% salt)
Production method: A 2 cm wide fillet of Fukuragagi was coated with the same weight of pickled bed, placed in a bag, and vacuum-packaged. As a general production method, fillets of Fukuragagi were wrapped in gauze, the same amount of seasoning lees as the ingredients was applied thereon, placed in a bag, vacuum-packaged, and stored at room temperature for 3 days.
Evaluation method glucose concentration: After homogenizing the fish meat after each treatment, 5 ml of 8% trichloroacetic acid aqueous solution was added to 5 g of the sample, vigorously shaken, filtered and clarified. The glucose concentration of the constant-volume liquid was measured using a glucose CII test manufactured by Wako Pure Chemical Industries, Ltd., and converted to the glucose concentration contained in the original sample. Three replicates were performed per treatment, and the mean and standard deviation were determined. As for the measurement results, as shown in FIG. 99, it was confirmed that 0.3% or more of glucose permeation was observed regardless of the treatment conditions compared to the untreated. The glucose concentration increases in proportion to the increase in treatment temperature. Also, the glucose concentration increases in proportion to the length of treatment time. Furthermore, the glucose concentration was higher than that in the general method at a treatment time of 15 hours at 40°C or higher, and at a treatment temperature of 40°C for 15 hours or longer.
Hardness: Each sample was prepared into a cube of 1 cm square, and a razor-blade plunger was applied perpendicularly to muscle fibers, and the stress was measured by a rheometer when the muscle fibers were penetrated to a depth of 8 mm. As for the measurement results, as shown in FIG. 100, it was observed that the pickled Fukuraguri lees tended to become harder when subjected to high-pressure heating than when untreated or by the general method. However, when treated for 15 hours, Fukuragi became softer in proportion to the increase in treatment temperature, and at 60°C, it became softer than untreated and conventional method. In addition, Fukuragaki softened in proportion to the length of treatment time.
Color tone: L*, a*, and b* were measured by the reflection method using a color difference meter (manufactured by Nippon Denshoku Co., Ltd.) for the color tone of the sample under each processing condition. As shown in FIG. 101, the measurement results show that the high-pressure heating process permeates the components and makes the material bright brown. Also, the vividness of the color is improved in proportion to the processing temperature and processing time. Also, FIG. 102(a) shows the color tone before the high-pressure heating treatment, and (b)
indicates the color tone before high-pressure heating treatment.
Microbial hygiene test method: Suspend each sample in physiological saline, serially dilute, and add each diluted sample to standard agar medium (Eiken, for general viable cell count) and 0.01 (w/v) % chloramphenicol. Potato dextrose agar medium (Eiken, for fungi) was smeared (in the case of standard agar, the pour method was used). After culturing at 30 to 37° C. for 3 days, the number of microbial colonies formed was counted, and the number of viable general bacteria and fungi per 1 g of sample was calculated. With reference to the standards of the Food Sanitation Act, the standard values were 300 cfu/g or less for the general viable count and 1000 cfu/g or less for the fungal count. Non-detection was defined as 50 cfu/g or less for both general viable bacteria and fungi.
(Table 47A) shows the results of microbial hygiene inspection by treatment temperature (all treatment times are 15 hours), and (Table 47B) shows the results of microbial hygiene inspections by treatment time (all treatment temperatures are 40°C).

In the table, above the standard means 100,000/g or more for general viable bacteria, 1000/g or more for fungi, and below the standard means 100,000/g or less for general viable bacteria, 1000/g or less for fungi, and no Detection refers to the limit of detection, 50/g or less.
As shown in the table, the viable cell counts of general viable bacteria and fungi were not detected or were below the sanitary standard values for both the treatment temperature and the treatment time. This is thought to be due not only to the effect of high-pressure heating, but also to the synergistic effect of the sterilization effect of the alcohol contained in the sake lees soaked in Fukuragagi, which inhibits the growth of bacteria, similar to pickled vegetables in sake lees.

(2) Squid pickled in sake lees/treatment conditions Pressure: 1000 atmospheres (100 MPa)
Temperature: 20°C to 60°C
Time: 7.5 hours to 30 hours Production details Seafood: Japanese flying squid (body)
Pickled bed: Seasoning lees (sake lees 73%, powdered rice koji 11.2%, sake 7.5%, mirin 7.5%, salt 0.8%)
Production method: The body of the peeled squid was coated with the same weight of pickling, placed in a bag, and vacuum-packaged. As a general production method, squid fillet was wrapped in gauze, the same amount of seasoning lees as the food was applied on top, put in a bag, vacuum-packaged, and stored at room temperature for 3 days.
Evaluation method glucose concentration: After homogenizing the squid meat after each treatment, 5 ml of 8% trichloroacetic acid aqueous solution was added to 5 g of the sample, vigorously shaken, filtered and clarified, and the volume of the liquid was adjusted to 50 ml. The glucose concentration of the constant-volume liquid was measured using a glucose CII test manufactured by Wako Pure Chemical Industries, Ltd., and converted to the glucose concentration contained in the original sample. Three replicates were performed per treatment, and the mean and standard deviation were determined. As shown in FIG. 103, the measurement results confirmed permeation of 0.5% or more of glucose, regardless of the treatment conditions, compared to the untreated. The glucose concentration increases in proportion to the increase in treatment temperature. Also, the glucose concentration increases in proportion to the length of treatment time. Furthermore, the concentration of glucose was higher than that in the general method regardless of the treatment conditions.
Hardness: Each sample was prepared into a cube of 2 cm square, and the stress was measured by a rheometer when a 5 mmφ needle-like plunger was used to penetrate the sample to 15 mm. As shown in FIG. 104, the squid pickled in sake lees also tended to become harder when subjected to high-pressure heating than when not treated, but was softer than when treated under any conditions. In the case of treatment for 15 hours, the treatment temperature was 40°C or more, and in the case of treatment at 40°C, the treatment time was 15 hours or more.
Color tone: L*, a*, and b* were measured by the reflection method using a color difference meter (manufactured by Nippon Denshoku Co., Ltd.) for the color tone of the sample under each processing condition. As for the measurement result, as shown in FIG. 105, by performing the high-pressure heating treatment, the components and the like permeate and become bright brown. Also, the vividness of the color is improved in proportion to the processing temperature and processing time. Also, FIG. 106(a) shows the color tone before high pressure heating treatment, and FIG. 106(b) shows the color tone before high pressure heating treatment.
Microbial hygiene test method: Suspend each sample in physiological saline, serially dilute, and add each diluted sample to standard agar medium (Eiken, for general viable cell count) and 0.01 (w/v) % chloramphenicol. Potato dextrose agar medium (Eiken, for fungi) was smeared (in the case of standard agar, the pour method was used). After culturing at 30 to 37° C. for 3 days, the number of microbial colonies formed was counted, and the number of viable general bacteria and fungi per 1 g of sample was calculated. With reference to the standards of the Food Sanitation Act, the standard values were 300 cfu/g or less for the general viable count and 1000 cfu/g or less for the fungal count. Non-detection was defined as 50 cfu/g or less for both general viable bacteria and fungi.
(Table 48A) shows the results of the microbial hygiene inspection by treatment temperature (all treatment times were 15 hours), and (Table 48B) shows the results of the microbial hygiene inspection by treatment time (all treatment temperatures were 40°C).

In the table, above the standard means 100,000/g or more for general viable bacteria, 1000/g or more for fungi, and below the standard means 100,000/g or less for general viable bacteria, 1000/g or less for fungi, and no Detection refers to the limit of detection, 50/g or less.
As shown in the table, the viable cell counts of general viable bacteria and fungi were not detected or were below the sanitary standard values for both treatment temperature and treatment time. It is thought that the multiplication of bacteria was suppressed not only by the effect of high-pressure heating, but also by the synergistic effect of the sterilization effect of the alcohol contained in the sake lees soaked in the squid, similar to pickling vegetables and Fukuragagi in sake lees.

(3) Fukurugi soft smoke/treatment conditions Pressure: 1000 atmospheres (100 MPa)
Temperature: 20°C to 60°C
Time: 30 minutes to 2 hours Production details Seafood: Fukuragi marinated liquid: Seasoning liquid (20% salt, 20% sugar, 8% trehalose, 2% powdered liquid, 50% water)
Production method: A 2 cm wide fillet of Fukuragagi and the same weight of pickling liquid were placed in a bag and vacuum-packaged.・Evaluation method Sugar concentration: A high-pressure heating-treated sample was taken out of the bag, and after wiping off the seasoning liquid, etc., it was homogenized with a mixer. After adding 1 ml of an 8% trichloroacetic acid solution to 1 g of the homogenized sample and stirring vigorously, the extract was filtered and diluted to 10 ml with distilled water to obtain a sample. An HPLC system (Shimadzu Corporation) was used for analysis. The column was Mightysil NH ₂ (5 μm) (Kanto Kagaku Co., Ltd.), the detector was a differential refractometer (RID-10A), the mobile phase was 70% acetonitrile, and the analysis was performed at a flow rate of 1 ml/min. Three components, fructose, glucose and sucrose, were evaluated by measuring the sugar composition of the sample and quantifying them. As shown in FIG. 107, the measurement results show that the high-pressure heating process increases the sugar concentration. No difference due to treatment temperature was observed.
Hardness: Each sample was prepared into a cube of 1 cm square, and a razor-blade plunger was applied perpendicularly to muscle fibers, and the stress was measured by a rheometer when the muscle fibers were penetrated to a depth of 8 mm. As shown in FIG. 108, the measurement results show that during the manufacturing process, dehydration occurs due to the salt content of the seasoning liquid, and drying treatment is performed. Effects of the high-pressure heating treatment include softening in proportion to the rise in treatment temperature and hardening in proportion to the length of treatment time.
Color tone: L*, a*, and b* were measured by the reflection method using a color difference meter (manufactured by Nippon Denshoku Co., Ltd.) for the color tone of the sample under each processing condition. As for the measurement result, as shown in FIG. 109, by performing the high-pressure heating process, the seasoning liquid permeates and becomes bright brown. Color vibrancy improves in proportion to treatment temperature and treatment time. FIG. 110(a) shows the color tone before the high pressure heating treatment, and FIG. 110(b) shows the color tone before the high pressure heating treatment.
Microbial hygiene test method: Suspend each sample in physiological saline, serially dilute, and add each diluted sample to standard agar medium (Eiken, for general viable cell count) and 0.01 (w/v) % chloramphenicol. Potato dextrose agar medium (Eiken, for fungi) was smeared (in the case of standard agar, the pour method was used). After culturing at 30 to 37° C. for 3 days, the number of microbial colonies formed was counted, and the number of viable general bacteria and fungi per 1 g of sample was calculated. With reference to the standards of the Food Sanitation Act, the standard values were 300 cfu/g or less for the general viable count and 1000 cfu/g or less for the fungal count. Non-detection was defined as 50 cfu/g or less for both general viable bacteria and fungi.
(Table 49A) shows the results of microbial hygiene inspection by treatment temperature (all treatment times are 1 hour), and (Table 49B) shows the results of microbial hygiene inspection by treatment time (all treatment temperatures are 40°C).

In the table, above the standard means 100,000/g or more for general viable bacteria, 1000/g or more for fungi, and below the standard means 100,000/g or less for general viable bacteria, 1000/g or less for fungi, and no Detection refers to the limit of detection, 50/g or less.
As shown in the table, by treatment temperature, neither general viable bacteria nor fungi were detected at treatment temperatures of 60°C or higher. By treatment time at 40℃, it was below the sanitary standard value for 2 hours or more.

(4) Soft smoked squid/treatment conditions Pressure: 1000 atmospheres (100 MPa)
Temperature: 20°C to 60°C
Time: 30 minutes to 2 hours Production details Seafood: Japanese flying squid (body)
Pickling liquid: Seasoning liquid (20% salt, 20% sugar, 8% trehalose, 2% powdered liquid, 50% water)
Production method: The skin-peeled squid body and the same weight of pickling liquid were placed in a bag and vacuum-packaged.
・Evaluation method Sugar concentration: A high-pressure heating-treated sample was taken out of the bag, and after wiping off the seasoning liquid, etc., it was homogenized with a mixer. After adding 1 ml of 8-trichloroacetic acid solution to 1 g of the homogenized sample and stirring vigorously, the extract was filtered and diluted to 10 ml with distilled water to obtain a sample. An HPLC system (Shimadzu Corporation) was used for analysis. The column was Mightysil NH ₂ (5 μm) (Kanto Kagaku Co., Ltd.), the detector was a differential refractometer (RID-10A), the mobile phase was 70% acetonitrile, and the analysis was performed at a flow rate of 1 ml/min. Three components, fructose, glucose, and sucrose, were evaluated by measuring the sugar composition of samples and quantifying them. As for the measurement results, as shown in FIG. 111, the high-pressure heating treatment increases the sugar concentration in the same manner as in Fukuraguri. Moreover, the sugar concentration increased the most when the treatment temperature was 60°C.
Hardness: Each sample was prepared into a cube of 2 cm square, and the stress was measured by a rheometer when it was penetrated to 15 mm by a needle-like plunger of 5 mmφ. As shown in FIG. 112, the measurement results show that, similarly to Fukuragi, it tends to become harder during the manufacturing process than untreated. Contrary to Fukuragi, the effect of high-pressure heating softens in proportion to the length of treatment time.
Color tone: L*, a*, and b* were measured by the reflection method using a color difference meter (manufactured by Nippon Denshoku Co., Ltd.) for the color tone of the sample under each processing condition. As for the measurement results, as shown in FIG. 113, by performing the high-pressure heating process, the seasoning liquid permeates and becomes bright brown like Fukuragagi. Also, the vividness of the color is improved in proportion to the treatment temperature and treatment time. FIG. 114(a) shows the color tone before the high pressure heating treatment, and (b) shows the color tone before the high pressure heating treatment.
Microbial hygiene test method: Suspend each sample in physiological saline, serially dilute, and add each diluted sample to standard agar medium (Eiken, for general viable cell count) and 0.01 (w/v) % chloramphenicol. Potato dextrose agar medium (Eiken, for fungi) was smeared (in the case of standard agar, the pour method was used). After culturing at 30 to 37° C. for 3 days, the number of microbial colonies formed was counted, and the number of viable general bacteria and fungi per 1 g of sample was calculated. With reference to the standards of the Food Sanitation Act, the standard values were 300 cfu/g or less for the general viable count and 1000 cfu/g or less for the fungal count. Non-detection was defined as 50 cfu/g or less for both general viable bacteria and fungi.
(Table 50A) shows the results of microbial hygiene inspection by treatment temperature (all treatment times are 1 hour), and (Table 50B) shows the results of microbial hygiene inspection by treatment time (all treatment temperatures are 40°C).

In the table, above the standard means 100,000/g or more for general viable bacteria, 1000/g or more for fungi, and below the standard means 100,000/g or less for general viable bacteria, 1000/g or less for fungi, and no Detection refers to the limit of detection, 50/g or less.
As shown in the table, neither general viable bacteria nor fungi were detected in soft smoked squid other than those treated at 40°C for 1 hour immediately after production. Soft smoked squid can be stored at 5°C for 1 month under any processing conditions.

(5) Effect of treatment at 80 MPa or less and 100 MPa or more When the method for producing pickled and soft smoked Fukuragi and squid was examined by changing only the pressure condition of the high pressure treatment condition in the range of 80 to 300 MPa, the results are shown in Table 51. , confirmed that the present invention is effective even in the pressure range of 80 to 300 MPa.

The degassing, heating, and high-pressure treatment method according to the present invention can be applied to vegetables other than the above in narazuke of vegetables, lightly pickled vegetables, and raw soy sauce pickles of vegetables. can be applied to other than apples, apricots, and plums. For pickled meat, it can be used not only for pork loin, but also for chicken and beef. It can also be used for general tofu, and for processed mushroom products, it can be used for mushrooms other than shiitake mushrooms.

Claims (1)

A degassing, heating, and high-pressure treatment method applied to the production of food, wherein the food is Japanese pears or grapes pickled in syrup, and the Japanese pears or grapes are placed in a container such as a polyethylene bag together with the syrup, and degassed. Only Japanese pears or grapes and syrup are held in the container, and the Japanese pears or grapes are placed in the container such as a polyethylene bag, and the pressure is 100 MPa and 40°C or higher and 70°C or lower for 15 minutes to 60 minutes. A degassing/heating/high-pressure treatment method characterized by performing a minute treatment.

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