CN110846229A - Preservation method and preservative for improving survival rate of bacillus coagulans - Google Patents
Preservation method and preservative for improving survival rate of bacillus coagulans Download PDFInfo
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- CN110846229A CN110846229A CN201911316054.8A CN201911316054A CN110846229A CN 110846229 A CN110846229 A CN 110846229A CN 201911316054 A CN201911316054 A CN 201911316054A CN 110846229 A CN110846229 A CN 110846229A
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Abstract
The invention discloses a preservation method and a preservative for improving the survival rate of bacillus coagulans. The preservation method comprises the step of mixing and preserving bacillus coagulans liquid with spores and a preservative, wherein the preservative comprises 0.35-1 g/mL of reducing agent, 20-40 g/mL of protective agent, 5-12 g/mL of cane sugar and 5-8 g/mL of filling agent, the stress resistance of bacillus coagulans is improved, and the preservation quality and preservation time limit of bacillus coagulans are effectively prolonged by matching with the preservative.
Description
Technical Field
The invention relates to the technical field of bacillus coagulans, and particularly relates to a preservation method and a preservative for improving the survival rate of bacillus coagulans.
Background
The probiotics play a role of probiotics by inhibiting pathogenic bacteria in the gastrointestinal tract and maintaining the ecological balance of flora, are safe and harmless, and are one of products with wider application.
The bacillus coagulans is a gram-positive bacterium, the cell body is rod-shaped, the two ends are blunt, the bacillus coagulans is single, paired or short-chain, flagellate-free, the spore is terminal, the bacillus coagulans is facultative anaerobic, and the bacillus coagulans can decompose saccharides to generate lactic acid. The optimal temperature for the growth of the thalli is 37-45 ℃, and the optimal pH value is 6.6-7.0. It not only has the probiotic effect of lactic acid bacteria and bifidobacteria, but also has the remarkable characteristics of bacillus: high-temp. resistance, acid resistance and high-temp. resistance to bile salt.
Because the bacillus coagulans has the common characteristics of lactic acid bacteria and bacillus and good safety, the bacillus coagulans is more and more widely applied to the livestock industry.
Freeze-drying is the most common process for producing probiotic starter cultures, but most probiotics suffer from different degrees of impairment of their normal physiological and metabolic activities after freezing and drying, resulting in a reduction or even loss of cell activity and fermentation viability.
In view of this, the invention is particularly proposed.
Disclosure of Invention
The invention aims to provide a preservation method and a preservative for improving the survival rate of bacillus coagulans. The preservation method preserves the bacillus coagulans with spores, and can effectively prolong the preservation quality and preservation time limit of the bacillus coagulans by matching with a preservative.
The invention is realized by the following steps:
in a first aspect, embodiments of the present invention provide a preservation method for improving survival rate of bacillus coagulans, which includes mixing bacillus coagulans with a preservative for preservation;
the bacillus coagulans is a bacterium liquid containing spores;
the preservative comprises the following components: 0.35-1 g/mL reducing agent, 5-8 g/mL filler, 5-12 g/mL sucrose, and 20-40 g/mL protective agent.
At present, the method for preserving the bacillus coagulans is to culture the bacillus coagulans on an agar plate for 24 to 48 hours, collect strains, directly mix the strains with glycerol with a certain concentration uniformly, and then put the strains into a refrigeration condition at the temperature of minus 20 ℃.
In the application, the bacillus coagulans is cultured into a bacterial liquid containing spores and then is preserved, and the preservation time limit of the bacillus coagulans in a low-temperature environment is prolonged by utilizing the stress resistance of the spores. Meanwhile, the preservative provided by the application is matched and adopted during preservation.
The reducing agent in the preservative can consume oxygen in the culture medium, so that the thalli are in an anaerobic state, and the oxidation of the thalli is reduced; the protective agent in the preservative has hydrophilicity, and can stabilize the configuration of cell components by the affinity of hydrogen and ionic bonds for water and cells, so as to prevent damage to cells due to freezing or continuous sublimation of water. While the bulking agent crystallizes upon slow freezing, thereby providing a structural support for the cells and not reacting with the intracellular active components.
In an alternative embodiment, the preservative comprises the following components: 0.45-0.65 g/mL reducing agent, 5-7 g/mL filler, 5-8 g/mL sucrose, and 30-40 g/mL protective agent. Under the component proportion, the preservation quality of the bacillus coagulans can be further prolonged, the physiological function and the metabolic activity of the bacillus coagulans after refrigeration are maintained, and the loss or reduction of the cell activity and the fermentation activity after refrigeration is effectively delayed.
In an alternative embodiment, the bulking agent comprises mannitol.
Preferably, the protective agent comprises glycerol and/or trehalose.
In alternative embodiments, the reducing agent is at least one of cysteine hydrochloride, vitamin D, sodium thiosulfate, and ethylenediaminetetraacetic acid.
In an alternative embodiment, the reducing agent is cysteine hydrochloride.
In an optional embodiment, the bacteria liquid and the preservative are mixed according to the following ratio of (0.5-1.5): 1 by volume.
In an alternative embodiment, the spore volume percentage of the spore-containing bacterial liquid is 70% to 90%.
In an alternative embodiment, the culture conditions for forming the spore-containing bacterial liquid are as follows: shaking-culturing at 37-40 deg.C and 180-200 r/min for 3-7 days.
In an optional embodiment, the culture medium of the bacterial liquid is an MRS liquid culture medium.
In alternative embodiments, the MRS liquid medium comprises: 10-14 g/L peptone, 10-14 g/L beef extract, 3-8 g/L yeast powder, 15-25 g/L glucose, 3-8 g/L sodium acetate, 1-3 g/L diammonium citrate, 0.5-1.5 g/LTween80 and 1-3 g/LK2HPO4、0.38～0.88g/LMgSO4.7H2O and 0.1-0.5 g/LMnSO4.H2O。
In an alternative embodiment, the bacterial liquid is prepared by centrifuging the spore-forming bacterial liquid.
In an optional embodiment, the centrifugation condition is 4000-6000 r/min, and the centrifugation is performed for 10-15 min at 4 ℃;
in an alternative embodiment, the bacterial liquid is a bacterial suspension prepared by centrifugally eluting a spore-forming bacterial liquid;
in an alternative embodiment, the elution conditions are: and (4) eluting the centrifuged bacterial sludge by using physiological saline with the volume of 45-55 times of the centrifuged bacterial sludge.
In a second aspect, the embodiments of the present invention further provide a preservative for improving the survival rate of bacillus coagulans, wherein the preservative comprises the following components: 0.35-1 g/mL reducing agent, 5-8 g/mL filler, 5-12 g/mL sucrose, and 20-40 g/mL protective agent. Specifically, the preservative is the same as that in the preservation method described in any of the above embodiments, and details thereof are not repeated herein.
The invention has the following beneficial effects:
the embodiment of the invention provides a preservation method and a preservative for improving the survival rate of bacillus coagulans. The preservation method comprises the steps of mixing and preserving bacillus coagulans liquid with spores with a preservative, improving the stress resistance of bacillus coagulans, and simultaneously, matching with the preservative, wherein a reducing agent in the preservative can consume oxygen in a culture medium, so that thalli are in an anaerobic state, and the oxidation effect of the thalli is reduced; the protective agent in the preservative has hydrophilicity, and can stabilize the configuration of cell components by the affinity of hydrogen and ionic bonds to water and cells so as to prevent damage to the cells due to freezing or continuous sublimation of water; and the filler can be crystallized when being frozen at a low speed, so that a support structure is provided for cells, the filler can not react with active components in the cells, and the preservation quality and the preservation time limit of the bacillus coagulans are effectively prolonged.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
The features and properties of the present invention are described in further detail below with reference to examples.
Example 1
A preservation method for improving the survival rate of bacillus coagulans, which comprises the following steps:
(1) preparing a bacillus coagulans bacterial suspension:
inoculating pure and rejuvenated bacillus coagulans into an MRS liquid culture medium (the composition is shown in Table 1), wherein the inoculation amount is 5%, carrying out shake culture at 40 ℃ and 200r/min for 5d, and the spore rate in the visual field of microscopic bacteria liquid reaches more than 80%, so that the state of the bacteria liquid can be preserved.
TABLE 1 MRS liquid Medium
Centrifuging the obtained bacterial liquid with a large amount of spores, wherein the centrifugation condition is 4000r/min, and centrifuging for 10min at 4 ℃; then, 0.85 percent of sterile normal saline with the volume 50 times of that of the bacterial sludge is adopted to elute the bacterial sludge, and the bacterial suspension of the bacillus coagulans is prepared.
(2) Preparation of a preservative:
the components of the preservative are shown in table 2.
TABLE 2 composition of preservative
| Components | (g/mL) |
| Glycerol | 40.00 |
| Sucrose | 5.00 |
| Mannitol | 6.00 |
| Cysteine hydrochloride | 0.50 |
(3) Mixing and preserving a bacillus coagulans bacterial suspension forming spores with a preservative:
mixing the bacterial suspension prepared in the step (1) and the preservative prepared in the step (2) according to a volume ratio of 1: 1, mixing, and then subpackaging in a freezing storage tube for preservation at-20 ℃.
Example 2
This example provides a preservation method for increasing the survival rate of bacillus coagulans, which is substantially the same as example 1 except for the following parameters:
(1) preparing a bacillus coagulans bacterial suspension:
inoculating the pure and rejuvenated bacillus coagulans into an MRS liquid culture medium (the composition is shown in Table 1), wherein the inoculation amount is 10%, carrying out shake culture at 40 ℃ and 200r/min for 3d, and the spore rate in the visual field of microscopic bacteria liquid reaches more than 70%, so that the state of the bacteria liquid can be preserved.
Example 3
This example provides a preservation method for increasing the survival rate of bacillus coagulans, which is substantially the same as example 1 except for the following parameters:
(1) preparing a bacillus coagulans bacterial suspension:
inoculating pure and rejuvenated bacillus coagulans into an MRS liquid culture medium (the composition is shown in Table 1), wherein the inoculation amount is 5%, carrying out shake culture at 40 ℃ and 200r/min for 9d, and the spore rate in the visual field of microscopic bacteria liquid reaches over 90%, so that the state of the bacteria liquid can be preserved.
Example 4
This example provides a preservation method for increasing the survival rate of bacillus coagulans, which is substantially the same as example 1 except that the preservative is different as follows:
the preservative agent comprises: 0.35g/mL cysteine hydrochloride, 20g/mL glycerol, 5g/mL mannitol and 5g/mL sucrose.
Example 5
This example provides a preservation method for increasing the survival rate of bacillus coagulans, which is substantially the same as example 1 except that the preservative is different as follows:
the preservative agent comprises: 1g/mL cysteine hydrochloride, 40g/mL glycerol, 8g/mL mannitol and 12g/mL sucrose.
Example 6
This example provides a preservation method, which is substantially the same as that of example 1, except that the culture medium is different from that of example 1, and the MRS medium is replaced with a YPD medium (tryptone 20.00g/L, yeast powder 10.00g/L, glucose 20.00g/L, agar powder 20.00g/L, pH 7.2).
Comparative example 1
This example provides a preservation method for increasing the survival rate of bacillus coagulans, which is substantially the same as example 1, except that the culture conditions for bacillus coagulans are different;
the culture conditions of the bacillus coagulans are as follows: culturing the bacillus coagulans on an agar plate for 24-48 h, and collecting the strain.
Comparative example 2
This example provides a preservation method for increasing the survival rate of bacillus coagulans, which is substantially the same as example 1, except that the preservative is prepared as follows: the preservative agent is glycerol.
Comparative example 3
This example provides a preservation method for enhancing the survival rate of bacillus coagulans, which is substantially the same as example 1, except that the preservative is prepared as follows:
the preservative agent is prepared by replacing glycerol with polyethylene glycol with the same mass.
Comparative example 4
This example provides a preservation method, substantially the same as example 1, except that the preservative is prepared as follows:
the preservative agent replaces cysteine hydrochloride with cysteine with the same mass.
Comparative example 5
This example provides a preservation method, substantially the same as example 1, except that the preservative is prepared as follows:
the preservative agent is prepared by replacing mannitol with glycerol in equal weight parts.
Verification example 1
The effect of the preservation methods provided herein on the survival rate of bacillus coagulans was verified.
The same bacillus coagulans (the strain is preserved in the second floor strain room of institute of animal health research institute of agriculture and forestry, hokkaido) is preserved by adopting the preservation methods provided in examples 1-6 and comparative examples 1-5.
Viable bacteria were counted on the refrigerated samples at day 1, day 30, day 60, day 90, day 120, day 150 and day 180, respectively, and the survival rate was calculated, and the results are shown in table 3.
TABLE 3 survival rate of Bacillus coagulans
As shown in Table 3, the experimental results obtained in examples 1 to 6 all prove that the combination of the preservative provided in the present invention for strain cryopreservation under the condition of high spore rate can effectively ensure that the viable bacteria amount and activity of the strain are not reduced for a long time.
Comparative examples 1 to 5 experiments comparing the present invention with different culture media, types and components of preservative agents, etc. have less preservation effect on strains than example 1. Therefore, the preservation method and the preservative provided by the invention can effectively keep the activity of the bacillus coagulans at low freezing temperature, wherein the best preservation effect is shown in example 1.
In summary, the embodiment of the invention provides a preservation method and a preservative for improving the survival rate of bacillus coagulans, the preservation method comprises the steps of mixing and preserving bacillus coagulans liquid with spores and the preservative, so that the stress resistance of bacillus coagulans is improved, and meanwhile, the preservative is used in a matched manner, and a reducing agent in the preservative can consume oxygen in a culture medium, so that thalli are in an anaerobic state, and the oxidation effect of the thalli is reduced; the protective agent in the preservative has hydrophilicity, and can stabilize the configuration of cell components by the affinity of hydrogen and ionic bonds to water and cells so as to prevent damage to the cells due to freezing or continuous sublimation of water; and the filler can be crystallized when being frozen at a low speed, so that a support structure is provided for cells, the filler can not react with active components in the cells, and the preservation quality and the preservation time limit of the bacillus coagulans are effectively prolonged.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. A preservation method for improving the survival rate of bacillus coagulans is characterized by comprising the steps of mixing bacillus coagulans with a preservative for preservation;
the bacillus coagulans is a bacterium liquid containing spores;
the preservative comprises the following components: 0.35-1 g/mL reducing agent, 5-8 g/mL filler, 5-12 g/mL sucrose, and 20-40 g/mL protective agent.
2. The preservation method for improving survival rate of bacillus coagulans according to claim 1, wherein the preservation agent comprises the following components: 0.45-0.65 g/mL reducing agent, 5-7 g/mL filler, 5-8 g/mL sucrose, and 30-40 g/mL protective agent.
3. The preservation method for improving survival rate of bacillus coagulans according to claim 1, wherein the bulking agent comprises mannitol;
preferably, the protective agent comprises glycerol and/or trehalose;
preferably, the reducing agent is at least one of cysteine hydrochloride, vitamin D, sodium thiosulfate and ethylenediamine tetraacetic acid;
preferably, the reducing agent is cysteine hydrochloride.
4. The preservation method for improving the survival rate of bacillus coagulans according to claim 1, wherein the ratio of the bacterial liquid to the preservative is (0.5-1.5): 1 by volume.
5. The preservation method for improving the survival rate of bacillus coagulans according to any one of claims 1 to 4, wherein the volume percentage of spores in the spore-containing bacterial liquid is 70-90%.
6. The preservation method for improving the survival rate of bacillus coagulans according to claim 5, wherein the culture conditions for forming the spore-containing bacterial solution are as follows: shaking-culturing at 37-40 ℃ and 180-200 r/min for 3-7 d.
7. The preservation method for improving the survival rate of bacillus coagulans according to claim 5, wherein the culture medium of the bacterial liquid is MRS liquid culture medium.
8. The preservation method for improving survival rate of bacillus coagulans according to claim 7, wherein the MRS liquid medium comprises: 10-14 g/L peptone, 10-14 g/L beef extract, 3-8 g/L yeast powder, 15-25 g/L glucose, 3-8 g/L sodium acetate, 1-3 g/L diammonium citrate, 0.5-1.5 g/LTween80 and 1-3 g/LK2HPO4、0.38～0.88g/LMgSO4.7H2O and 0.1-0.5 g/LMnSO4.H2O。
9. The preservation method for improving the survival rate of bacillus coagulans as claimed in claim 5, wherein the bacterial liquid is prepared by centrifuging a spore-forming bacterial liquid;
preferably, the centrifugation condition is 4000-6000 r/min, and the centrifugation is carried out for 10-15 min at 4 ℃;
preferably, the bacterial liquid is a bacterial suspension prepared by centrifugally eluting the bacterial liquid forming spores;
preferably, the elution conditions are: and (4) eluting the centrifuged bacterial sludge by using physiological saline with the volume of 45-55 times of the centrifuged bacterial sludge.
10. A preservative for improving the survival rate of bacillus coagulans, wherein the preservative comprises the following components: 0.35-1 g/mL reducing agent, 5-8 g/mL filler, 5-12 g/mL sucrose, and 20-40 g/mL protective agent;
preferably, the preservative comprises the following components: 0.45-0.65 g/mL of reducing agent, 5-7 g/mL of filling agent, 5-8 g/mL of cane sugar and 30-40 g/mL of protective agent;
preferably, the bulking agent comprises mannitol;
preferably, the protective agent comprises glycerol and/or trehalose;
preferably, the reducing agent is at least one of cysteine hydrochloride, vitamin D, sodium thiosulfate and ethylenediamine tetraacetic acid;
preferably, the reducing agent is cysteine hydrochloride.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113005052A (en) * | 2021-01-27 | 2021-06-22 | 武汉微康益生菌研究院有限公司 | Preservation method of bacillus coagulans BC99 |
| CN113151066A (en) * | 2021-03-31 | 2021-07-23 | 长沙和光生物科技有限公司 | Bacillus coagulans HALO178 and preparation method thereof |
| CN114854593A (en) * | 2022-03-31 | 2022-08-05 | 浙江泰林生命科学有限公司 | Suspension type biological indicator protective agent |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1427886A (en) * | 2000-05-02 | 2003-07-02 | 表飞鸣制药株式会社 | Spray-dried microbial cells |
| CN101173228A (en) * | 2006-11-01 | 2008-05-07 | 信谊药厂 | Novel lactic acid bacillus mycopremna, pharmaceutical composition and uses thereof, and production method |
| CN102028954A (en) * | 2009-09-29 | 2011-04-27 | 成都康弘生物科技有限公司 | Preparation of recombinant adenovirus |
| CN104480052A (en) * | 2015-01-05 | 2015-04-01 | 江苏省苏微微生物研究有限公司 | Composite bacillus preparation containing three strains, preparation method of composite bacillus preparation and application of composite bacillus preparation to ecological breeding |
| CN105779384A (en) * | 2016-03-30 | 2016-07-20 | 王凌仪 | Seed cell screening and culturing cryopreservation technical method of human amniotic mesenchymal stem cells for tissue engineering |
| CN106047769A (en) * | 2016-08-04 | 2016-10-26 | 北京好实沃生物技术有限公司 | Bacillus coagulans-containing composite inocula and preparation method thereof |
| CN109294916A (en) * | 2018-11-06 | 2019-02-01 | 哈尔滨艾诺卫康科技发展有限公司 | A kind of culture presevation agent and the method using the preserving agent anaerobism freezing genetic engineering bacterium |
| CN110387330A (en) * | 2019-07-17 | 2019-10-29 | 上海理工大学 | The freeze-drying method of lactobacillus plantarum survival rate is improved using composite protectant |
| US20190328798A1 (en) * | 2018-04-30 | 2019-10-31 | Muhammed Majeed | Modulation of immune function by bacillus coagulans |
-
2019
- 2019-12-19 CN CN201911316054.8A patent/CN110846229A/en not_active Withdrawn
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1427886A (en) * | 2000-05-02 | 2003-07-02 | 表飞鸣制药株式会社 | Spray-dried microbial cells |
| CN101173228A (en) * | 2006-11-01 | 2008-05-07 | 信谊药厂 | Novel lactic acid bacillus mycopremna, pharmaceutical composition and uses thereof, and production method |
| CN102028954A (en) * | 2009-09-29 | 2011-04-27 | 成都康弘生物科技有限公司 | Preparation of recombinant adenovirus |
| CN104480052A (en) * | 2015-01-05 | 2015-04-01 | 江苏省苏微微生物研究有限公司 | Composite bacillus preparation containing three strains, preparation method of composite bacillus preparation and application of composite bacillus preparation to ecological breeding |
| CN105779384A (en) * | 2016-03-30 | 2016-07-20 | 王凌仪 | Seed cell screening and culturing cryopreservation technical method of human amniotic mesenchymal stem cells for tissue engineering |
| CN106047769A (en) * | 2016-08-04 | 2016-10-26 | 北京好实沃生物技术有限公司 | Bacillus coagulans-containing composite inocula and preparation method thereof |
| US20190328798A1 (en) * | 2018-04-30 | 2019-10-31 | Muhammed Majeed | Modulation of immune function by bacillus coagulans |
| CN109294916A (en) * | 2018-11-06 | 2019-02-01 | 哈尔滨艾诺卫康科技发展有限公司 | A kind of culture presevation agent and the method using the preserving agent anaerobism freezing genetic engineering bacterium |
| CN110387330A (en) * | 2019-07-17 | 2019-10-29 | 上海理工大学 | The freeze-drying method of lactobacillus plantarum survival rate is improved using composite protectant |
Non-Patent Citations (3)
| Title |
|---|
| 徐文斌: "乳酸菌芽孢的发酵、保藏条件研究及其合生素在肉鸡饲料中的应用", 《中国优秀硕士学位论文全文数据库 农业科技辑》 * |
| 曾小兰: "《食品微生物及其检验技术》", 31 August 2010 * |
| 王世宇: "《药用辅料学 十三五规划》", 30 April 2019 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113005052A (en) * | 2021-01-27 | 2021-06-22 | 武汉微康益生菌研究院有限公司 | Preservation method of bacillus coagulans BC99 |
| CN113151066A (en) * | 2021-03-31 | 2021-07-23 | 长沙和光生物科技有限公司 | Bacillus coagulans HALO178 and preparation method thereof |
| CN113151066B (en) * | 2021-03-31 | 2023-02-07 | 长沙和光生物科技有限公司 | Bacillus coagulans HALO178 and preparation method thereof |
| CN114854593A (en) * | 2022-03-31 | 2022-08-05 | 浙江泰林生命科学有限公司 | Suspension type biological indicator protective agent |
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