JP2005333851A - Fermentation promotor - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an entirely new composition containing a nitrogen source, a carbon source, vitamins, a mineral, phosphorus and the like, and derived from rice bran by extracting components contained in the rice bran as an essence. <P>SOLUTION: The fermentation promotor contains the components extracted from the rice bran and containing the mineral, the water-soluble vitamins, glucide and phytic acid as active ingredients. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

    The present invention is a composition excellent in the fermentation-promoting effect derived from rice bran for the purpose of promoting and promoting the growth of microorganisms in the field of using microorganisms such as fermented foods, food additives by fermentation technology and production of substances such as pharmaceuticals. Is to provide.

  Conventionally, natural components such as peptone, malt extract, and meat extract have been used as sources of nitrogen and trace components to promote growth when culturing microorganisms. As these raw materials, fish meat, animal meat, milk, soybeans, wheat and the like are used, and they are used in fields using microorganisms such as fermented foods. In addition, the fermentation is usually promoted by adding vitamins or minerals.

On the other hand, rice bran has been used as a mushroom culture medium. In addition, since long ago, studies have been conducted on using rice bran as a fertilizer or food material by fermenting rice bran with specific microorganisms (see, for example, Patent Document 1). This indicates that rice bran contains nutrients necessary for the growth of microorganisms. However, rice bran itself has a high content of insoluble dietary fiber that is difficult for microorganisms to use and is insoluble, so it is not suitable for liquid culture and contains a large amount of oil, which adversely affects the culture system. For these reasons, it is impossible to use it for fermentation as an auxiliary for the purpose of promoting the growth of microorganisms.
JP 2004-33134 A

  The object of the present invention is to extract a component contained in rice bran as an extract to obtain a composition having a completely novel fermentation-promoting effect derived from rice bran, including a nitrogen source, a carbon source, vitamins, minerals, phosphorus and the like. Is the purpose.

  As a result of intensive efforts to solve the above-mentioned problems, the present inventors have obtained a fermentation promoter comprising as an active ingredient a rice bran extract component containing minerals, water-soluble vitamins, carbohydrates and phytic acid in rice bran.

  Unprecedented ability to obtain a complex fermentation promoter that contains nitrogen source, carbon source, phosphorus, vitamins, and minerals using rice bran as a raw material, and production of substances such as fermented foods and food additives and pharmaceuticals by fermentation technology In addition, it can be used as a fermentation promoter that promotes and promotes the growth of microorganisms in various fields that utilize microorganisms.

  In the present invention, in order to achieve the above object, when performing extraction from rice bran or defatted rice bran, an extract is obtained by using an organic acid contained in rice bran, in particular, an extract containing phytic acid, and dried. Manufactures rice bran extract powder. Here, phytic acid is an ingredient originally contained in rice bran, and there is no problem due to inclusion in rice bran extract. The other organic acid may be any organic acid that is recognized as an optional food additive when the fermentation product is used as a food. For example, acetic acid, oxalic acid, tartaric acid, phosphoric acid, lactic acid, butyric acid, Examples include citric acid and succinic acid. Thus, by specifying the kind of organic acid, the process of removing these acids is not necessary, and the solubility of the powder obtained by drying the acid can be maintained. In particular, when an extract containing phytic acid is used for extraction, since the extract obtained contains no components other than rice bran-derived components, an extract from pure rice bran can be obtained.

  Further, even if the extraction solution is extracted with water or an appropriate aqueous solution of salt or alkali, the composition intended by the present invention can be obtained.

  If the pH at the time of extraction is about 5.5 or less, a considerable amount of phytin can be extracted and the object of the present invention can be achieved. However, in consideration of the corrosiveness to the production apparatus, the pH at the time of extraction is In the range of 4.5 to 5.5. About 30 minutes is sufficient for the extraction time. In addition, depending on the target fermentation form, the extract is not limited to this pH, and an extract can be obtained using each of the aforementioned aqueous solutions. Then, the extract is separated and recovered from the defatted lees by a method such as centrifugation. The obtained extract is a yellowish white opaque liquid.

  The obtained extract may be used as it is, or a membrane treatment such as ultrafiltration or a purification treatment such as precipitation fraction is appropriately performed.

  Furthermore, the rice bran extract powder which is the rice bran extract which concerns on this invention is obtained by drying an extract. As a drying method, it is desirable to use a spray dryer or a freeze dryer in order to suppress the denaturation of the rice bran extract component as much as possible, but other methods may be used as long as the drying method can suppress the denaturation. In addition, it is desirable to concentrate the extract in advance for efficient drying.

  The rice bran extract used in the present invention is, for example, protein: 8.2 g / 100 g, carbohydrate: 57.7 g / 100 g (including sucrose 25.5 g / 100 g, dietary fiber: 11.1 g / 100 g), Potassium: 5.33 g / 100 g, Calcium: 121 mg / 100 g, Magnesium: 2.65 g / 100 g, physiologically active IP6: 27.8 g / 100 g, vitamin B1: 7.19 mg / 100 g, vitamin B2: 0.63 mg / 100 g, vitamin B6: 10.2 mg / 100 g, iron: 19.1 mg / 100 g, zinc: 13.2 mg / 100 g, inositol: 11.1 g / 100 g.

  The rice bran extract thus obtained is in a suitable form such as powder or aqueous solution depending on the intended use as a conventional fermentation accelerator, or as a fermentation accelerator with the supply of micronutrients, sugars and phosphorus in mind. Used in Purpose as a fermented food preparation or a fermented food that is flexible and safe for the production of substances such as food additives, pharmaceuticals, etc., or a fermented food with the aim of supplying micronutrients, dietary fiber, phosphorus and protein Can be used. Fields used include sake brewing such as sake, beer, fruit liquor, whiskey and shochu, other brewing fields such as soy sauce, miso and vinegar, dairy products and bread including yogurt, cheese and whey beverages. Fermented foods such as natto, organic acid fermentation, amino acid and nucleic acid fermentation and other low molecular substance production fields, antibiotics and other inhibitors, enzyme and other biopolymer material production fields, and the use of microorganisms as a source of fats and proteins However, the present invention is not limited to these as long as it uses microorganisms, and any field may be used. Moreover, the addition amount of the rice bran extract to a general culture system is usually 0.01 to 50% by weight, preferably about 0.1 to 2% by weight. The rice bran extract component is not toxic and is excellent in water solubility, so that no particular problem occurs even if it is added in a large amount. Even when it is added as a supplement for minerals and water-soluble vitamins, it is contained in this component itself without adding any other nitrogen source or phosphorus source, so that it shows an effect on the growth of microorganisms. Next, the present invention will be specifically described with reference to examples.

  EXAMPLES Hereinafter, although an Example is given and this invention is demonstrated further in detail, this invention is not limited to these.

  The pH was adjusted to 5.0 by adding 200 liters of water to 20 kg of defatted lees and adding a 50% aqueous phytic acid solution. This was stirred for 30 minutes and extracted, and then the cocoon and the extract were separated by a centrifuge. After the extract was concentrated to 50 liters, the concentrate was dried with a spray dryer to obtain a rice bran extract as 5.3 kg of rice bran extract powder. Table 1 shows the content of each component of the rice bran extract.

  As shown in Table 1, the rice bran extract component contains a large amount of phytic acid, which is a phosphoric acid compound present in rice bran, in addition to the rice bran-derived inorganic substance, vitamin B group, saccharides, and protein. Examples relating to these characteristics will be described below.

To M9 basic medium (disodium hydrogen phosphate: 6 g / L, potassium dihydrogen phosphate: 3 g / L, sodium chloride: 5 g / L, ammonium chloride: 1 g / L, magnesium sulfate: 1 mM, thiamine chloride 4 mg / L) A 1% rice bran extract according to the present invention was added, and the mixture was filtered through a cellulose acetate filter unit (0.2 μm) to prepare a medium.
5 ml of the medium thus prepared was put into a test tube sterilized by dry heat, washed with sterilized physiological saline pre-cultured with Bacillus Natto, and 0.1 ml of a suspension suspended at an equal magnification. After inoculation, shaking culture was performed at 30 ° C. (shaking frequency: 150 reciprocations / minute, test tube angle: 45 °), and the cell growth was measured for turbidity at 600 nm (OD600).
As a comparison of rice bran extract, 1% glucose, tryptone (BACTO), yeast extract (BACTO), and malt extract (BACTO) were added to the rice bran extract and cultured in the same manner.
As a result, as shown in FIG. 1, when the rice bran extract was added, the growth was 6 times that of glucose and 2 times that of the malt extract after 22 hours of culture, and the growth was similar to that of the yeast extract and tryptone. From this, since the promotion effect with respect to the growth of the natto bacteria of the rice bran extract was confirmed, the fermentation promotion effect in the said bacteria is clear.

To M9 basic medium (disodium hydrogen phosphate: 6 g / L, potassium dihydrogen phosphate: 3 g / L, sodium chloride: 5 g / L, ammonium chloride: 1 g / L, magnesium sulfate: 1 mM, thiamine chloride 4 mg / L) A 1% rice bran extract according to the present invention was added, and the mixture was filtered through a cellulose acetate filter unit (0.2 μm) to prepare a medium.
5 ml of the medium thus prepared is put into a test tube sterilized by dry heat, and yeast (Saccharomyces cerevisiae) pre-cultured with a complete medium is washed with sterilized physiological saline, and then 0.1 ml of a 1 × suspension is implanted. After the fungus, shaking culture was performed at 30 ° C. (shaking frequency: 150 reciprocations / minute, test tube angle: 45 °), and the cell growth was measured for turbidity at 600 nm (OD600).
As a comparison with rice bran extract, 1% glucose, tryptone (BACTO), yeast extract (BACTO), and malt extract (BACTO) were added to the rice bran extract and cultured in the same manner.
As a result, as shown in FIG. 2, the addition of rice bran extract showed 10 to 4 times the growth after 10 hours of culturing as compared to the other natural ingredient added groups, and each natural component was 4 to 4 times after 22 hours of culturing. The growth was twice as much. From this, since the promotion effect with respect to the proliferation of the yeast (Saccharomyces cerevisiae) of the rice bran extract was confirmed, the fermentation promotion effect in the said microbe is clear.

To M9 basic medium (disodium hydrogen phosphate: 6 g / L, potassium dihydrogen phosphate: 3 g / L, sodium chloride: 5 g / L, ammonium chloride: 1 g / L, magnesium sulfate: 1 mM, thiamine chloride 4 mg / L) A 1% rice bran extract according to the present invention was added, and the mixture was filtered through a cellulose acetate filter unit (0.2 μm) to prepare a medium.
5 ml of the medium thus prepared is put into a test tube sterilized by dry heat, and after washing yeast (Candida tropicalis) pre-cultured with a complete medium with sterilized physiological saline, 0.1 ml of a 1 × suspension is implanted. After the fungus, shaking culture was performed at 30 ° C. (shaking frequency: 150 reciprocations / minute, test tube angle: 45 °), and the cell growth was measured for turbidity at 600 nm (OD600).
As a comparison with rice bran extract, 1% glucose, tryptone (BACTO), yeast extract (BACTO), and malt extract (BACTO) were added to the rice bran extract and cultured in the same manner.
As a result, as shown in FIG. 3, when the rice bran extract was added, the growth was about 7 times that of glucose and about 2 times that of tryptone after 22 hours of culture, and the growth was equivalent to that of yeast extract and malt extract. From this, since the promotion effect with respect to the proliferation of the yeast of a rice bran extract was confirmed, the fermentation promotion effect in the said microbe is clear.

To M9 basic medium (disodium hydrogen phosphate: 6 g / L, potassium dihydrogen phosphate: 3 g / L, sodium chloride: 5 g / L, ammonium chloride: 1 g / L, magnesium sulfate: 1 mM, thiamine chloride 4 mg / L) A 1% rice bran extract according to the present invention was added, and the mixture was filtered through a cellulose acetate filter unit (0.2 μm) to prepare a medium.
5 ml of the medium thus prepared is put into a test tube sterilized by dry heat, washed with sterilized physiological saline preliminarily cultured in a complete medium, and then 0.1 ml of a 1 × suspension is implanted. After the fungus, shaking culture was performed at 30 ° C. (shaking frequency: 150 reciprocations / minute, test tube angle: 45 °), and the microbial cell growth was measured for turbidity at 600 nm (OD600).
As a comparison with rice bran extract, 1% glucose, tryptone (BACTO), yeast extract (BACTO), and malt extract (BACTO) were added to the rice bran extract and cultured in the same manner.
As a result, as shown in FIG. 4, the addition of rice bran extract showed 5 times the growth of glucose, 10 times the tryptone, and 5 times the malt extract after 22 hours of culture, and 70% of the yeast extract. Showed growth. From this, since the promotion effect with respect to the proliferation of the lactic acid bacteria (Lactbacillus casei) of the rice bran extract was confirmed, the fermentation promotion effect in the said microbe is clear.

To M9 basic medium (disodium hydrogen phosphate: 6 g / L, potassium dihydrogen phosphate: 3 g / L, sodium chloride: 5 g / L, ammonium chloride: 1 g / L, magnesium sulfate: 1 mM, thiamine chloride 4 mg / L) A 1% rice bran extract according to the present invention was added, and the mixture was filtered through a cellulose acetate filter unit (0.2 μm) to prepare a medium.
5 ml of the medium thus prepared was put into a test tube sterilized by dry heat and inoculated with Aspergillus niger spores pre-cultured in a complete medium, followed by shaking culture at 30 ° C. (shaking frequency: 150 reciprocations / minute) The angle of the test tube was 45 °), and the growth of the bacterial cells was measured by wet filtration after the culture solution was suction filtered through a membrane filter (5.0 μm).
As a comparison with rice bran extract, 1% glucose, tryptone (BACTO), yeast extract (BACTO), and malt extract (BACTO) were added to the rice bran extract and cultured in the same manner.
As a result, as shown in FIG. 5, the addition of the rice bran extract showed growth 4 times that of glucose, 10 times that of tryptone, and about 2 times that of malt extract and yeast extract after 48 hours of culture. From this, the promotion effect on the growth of Aspergillus niger by the rice bran extract was confirmed, and the fermentation promotion effect in the bacterium is clear.

To M9 basic medium (disodium hydrogen phosphate: 6 g / L, potassium dihydrogen phosphate: 3 g / L, sodium chloride: 5 g / L, ammonium chloride: 1 g / L, magnesium sulfate: 1 mM, thiamine chloride 4 mg / L) A 1% rice bran extract according to the present invention was added, and the mixture was filtered through a cellulose acetate filter unit (0.2 μm) to prepare a medium.
5 ml of the medium thus prepared was put into a test tube sterilized by dry heat and inoculated with spores of Aspergillus oyzae pre-cultured with a complete medium, followed by shaking culture at 30 ° C. (shaking frequency 150 reciprocations / minute) The angle of the test tube was 45 °), and the growth of the bacterial cells was measured by wet filtration after the culture solution was suction filtered through a membrane filter (5.0 μm).
As a comparison with rice bran extract, 1% glucose, tryptone (BACTO), yeast extract (BACTO), and malt extract (BACTO) were added to the rice bran extract and cultured in the same manner.
As a result, as shown in FIG. 6, when the rice bran extract was added, the growth was 10 times that of glucose, 2 times that of tryptone, and about 1.5 times that of the malt extract after 48 hours of culture, and the same growth as that of the yeast extract. showed that. From this, since the promotion effect with respect to the proliferation of the Aspergillus oyzae of the rice bran extract was confirmed, the fermentation promotion effect in the said microbe is clear.

To M9 basic medium (disodium hydrogen phosphate: 6 g / L, potassium dihydrogen phosphate: 3 g / L, sodium chloride: 5 g / L, ammonium chloride: 1 g / L, magnesium sulfate: 1 mM, thiamine chloride 4 mg / L) A 1% rice bran extract according to the present invention was added, and the mixture was filtered through a cellulose acetate filter unit (0.2 μm) to prepare a medium.
5 ml of the thus prepared medium is put into a test tube sterilized by dry heat, and 0.1 ml of a suspension in an equal volume is washed after washing with Bacillus subtilis pre-cultured with a complete medium with sterilized physiological saline. After inoculation, shaking culture was performed at 30 ° C. (shaking frequency: 150 reciprocations / minute, test tube angle: 45 °), and the cell growth was measured for turbidity at 600 nm (OD600).
As a comparison of rice bran extract, 1% glucose, tryptone (BACTO), yeast extract (BACTO), and malt extract (BACTO) were added to the rice bran extract and cultured in the same manner.
As a result, as shown in FIG. 7, the addition of the rice bran extract showed about 8 times as much glucose as that of the malt extract after 10 hours of culture, and about 50% of the growth of tryptone and yeast extract. From this, since the promotion effect with respect to the proliferation of the Bacillus subtilis of a rice bran extract was confirmed, the fermentation promotion effect in the said microbe is clear.

To M9 basic medium (disodium hydrogen phosphate: 6 g / L, potassium dihydrogen phosphate: 3 g / L, sodium chloride: 5 g / L, ammonium chloride: 1 g / L, magnesium sulfate: 1 mM, thiamine chloride 4 mg / L) A 1% rice bran extract according to the present invention was added, and the mixture was filtered through a cellulose acetate filter unit (0.2 μm) to prepare a medium.
5 ml of the medium thus prepared is put into a test tube sterilized by dry heat, and Escherichia coli pre-cultured with a complete medium is washed with sterilized physiological saline, and then 0.1 ml of a 1 × suspension is implanted. After the fungus, shaking culture was performed at 30 ° C. (shaking frequency: 150 reciprocations / minute, test tube angle: 45 °), and the cell growth was measured for turbidity at 600 nm (OD600).
As a comparison with rice bran extract, 1% glucose, tryptone (BACTO), yeast extract (BACTO), and malt extract (BACTO) were added to the rice bran extract and cultured in the same manner.
As a result, as shown in FIG. 7, the addition of the rice bran extract showed growth exceeding glucose after 10 hours of culture, and showed growth of about 60 to 70% of tryptone, yeast extract and malt extract. From this, since the promotion effect with respect to the proliferation of Escherichia coli of the rice bran extract was confirmed, the fermentation promotion effect in the said microbe is clear.

It is drawing which shows the addition effect of the rice bran extract with respect to growth of a microbial cell in culture | cultivation of Bacillus natto. It is drawing which shows the addition effect of the rice bran extract with respect to growth of a microbial cell in culture | cultivation of yeast (Saccharomyces cerevisiae). It is drawing which shows the addition effect of the rice bran extract with respect to growth of a microbial cell in culture | cultivation of yeast (Candida tropicalis). It is drawing which shows the addition effect of the rice bran extract with respect to growth of a microbial cell in culture | cultivation of lactic acid bacteria (Lactbacillus casei). It is drawing which shows the addition effect of the rice bran extract with respect to the growth of a microbial cell in culture | cultivation of a black mold (Aspergillus niger). It is drawing which shows the addition effect of the rice bran extract with respect to growth of a microbial cell in culture | cultivation of Aspergillus oryzae. It is drawing which shows the addition effect of the rice bran extract with respect to the growth of a microbial cell in culture | cultivation of Bacillus subtilis. It is drawing which shows the addition effect of the rice bran extract with respect to cell growth in culture | cultivation of colon_bacillus | E._coli (Escherichia coli).

Explanation of symbols

The ● marks in all the drawings indicate the amount of microorganisms in the medium in which the rice bran extract according to the present invention is added to the M9 basic medium. Similarly, ♦ indicates glucose, ■ indicates tryptone, ▲ indicates yeast extract, and * indicates the amount of microorganisms in the medium supplemented with malt extract.

Claims (4)

  A fermentation promoter comprising a rice bran extract containing minerals, water-soluble vitamins, sugars, phytic acid and protein as an active ingredient. The fermentation promoter according to claim 1, wherein the rice bran extract is a water-soluble extract of rice bran extracted from natural rice bran.   The extract according to claim 1 is selected from the group consisting of organic acids, acetic acid, oxalic acid, tartaric acid, phosphoric acid, lactic acid, butyric acid, citric acid, anhydrous citric acid, glacial acetic acid, benzoic acid and succinic acid contained in rice bran. Fermentation promoter that is extracted from one or more types. A fermentation promoter wherein the extract according to claim 1 is extracted with an extract containing water-soluble phytic acid.

Images