Screening method of bacillus coagulans capable of easily forming spores

A technology of Bacillus coagulans and a screening method, which is applied in the field of microorganisms, can solve the problems of unpopularity, poor repeatability, and complex sporulation mechanism, etc., and achieve the effect of improving genetic stability, simple method, and improving ability

Pending Publication Date: 2021-08-06
YATAI XINGYUAN FARMING TECH HAIAN CO LTD +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The high stress resistance of Bacillus coagulans comes from its ability to form spores, but the spore production ability of Bacillus coagulans is far inferior to that of commonly used Bacillus such as Bacillus subtilis and Bacillus licheniformis, and its spore production mechanism is more complicated
[0004] At present, most of the production of Bacillus coagulans is to increase the spore rate of Bacillus coagulans through the medium formula of rejuvenating components and complex feeding in the process, which leads to a fixed fermentation process, high requirements for equipment and raw material grades, and poor repeatability. , and cannot be popularized; moreover, some commonly used domestication methods for increasing the spore rate of spores are not suitable for Bacillus coagulans, and cannot be domesticated to obtain high-yield and high-spore rate Bacillus coagulans thallus

Method used

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  • Screening method of bacillus coagulans capable of easily forming spores
  • Screening method of bacillus coagulans capable of easily forming spores
  • Screening method of bacillus coagulans capable of easily forming spores

Examples

Experimental program
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Effect test

Embodiment 1

[0068] This embodiment provides a screening method for Bacillus coagulans that is prone to spore formation, which comprises the following steps:

[0069] (1) Strain rejuvenation

[0070] Inoculate the target Bacillus coagulans stored on the slant into a 1L conventional shake flask with a liquid volume of 200mL, culture the shake flask at 39-41°C and 200r / min for 28 hours, streak on the first solid plate, and store at 40°C Aerobic cultivation for 45 hours.

[0071] The formula of the shake flask medium is: peptone 10g / L, yeast extract powder 5g / L, glucose 10g / L, sodium acetate 5g / L, diammonium citrate 2g / L and dipotassium hydrogen phosphate 2g / L. The pH is 6.8.

[0072] Wherein the first solid plate adopts MRS agar.

[0073] (2) Baffle shake flask culture and water bath

[0074] Pick the dominant colony from the first solid plate and inoculate it into a 1L baffle shake flask with a liquid volume of 100mL, and culture the shake flask at 39-41°C and 200r / min for 42 hours, the...

Embodiment 2

[0080] The present embodiment provides a screening method for Bacillus coagulans which is easy to form spores, which comprises the following steps:

[0081] (1) Strain rejuvenation

[0082] Inoculate the target Bacillus coagulans stored on the slant into a 1L conventional shake flask with a liquid volume of 200mL, and culture the shake flask at 39-41°C and 180r / min for 24 hours, streak on the first solid plate, and keep at 39°C Aerobic cultivation for 42 hours.

[0083] The formula of shake flask medium is: peptone 8g / L, yeast extract powder 4g / L, glucose 8g / L, sodium acetate 4g / L, diammonium citrate 1.6g / L and dipotassium hydrogen phosphate 1.6g / L, The starting pH was 6.7.

[0084]Wherein the first solid plate adopts MRS agar.

[0085] (2) Baffle shake flask culture and water bath

[0086] Pick the dominant colony from the first solid plate and inoculate it into a 1L baffle shake flask with a liquid volume of 100mL, and culture the shake flask at 39-41°C and 180r / min for ...

Embodiment 3

[0092] This embodiment provides a screening method for Bacillus coagulans that is prone to spore formation, which comprises the following steps:

[0093] (1) Strain rejuvenation

[0094] Inoculate the target Bacillus coagulans stored on the slant into a 1L conventional shake flask with a liquid volume of 200mL, culture the shake flask at 39-41°C and 220r / min for 30 hours, streak on the first solid plate, and keep at 41°C Aerobic cultivation for 48 hours.

[0095] The formula of shake flask medium is: peptone 9g / L, yeast extract powder 4.5g / L, glucose 9g / L, sodium acetate 4.5g / L, diammonium citrate 1.8g / L and dipotassium hydrogen phosphate 1.8g / L L, with an initial pH of 6.9.

[0096] Wherein the first solid plate adopts MRS agar.

[0097] (2) Baffle shake flask culture and water bath

[0098] Pick the dominant colony from the first solid plate and inoculate it into a 1L baffle shake flask with a liquid volume of 100mL, and culture the shake flask at 39-41°C and 220r / min fo...

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Abstract

The invention discloses a screening method of bacillus coagulans capable of easily forming spores, and belongs to the technical field of microorganisms. The screening method comprises the following steps: repeatedly carrying out water bath-plate screening on rejuvenated bacillus coagulans for multiple times until the spore rate of the bacillus coagulans is not lower than 95%. According to the method, the rejuvenated bacillus coagulans are subjected to water bath method screening to produce spore individuals; then, the individuals, which do not form spores, are screened out through solid plate culture; multiple times of water bath-plate screening are repeated to improve the hereditary stability; the bacillus coagulans thalli, which easily form spores, are finally obtained through screening; and the spore rate after fermentation is improved while the yield is ensured. The method is simple, convenient to operate, capable of improving the spore forming capacity of the bacillus coagulans, not prone to strain degradation, capable of greatly improving the survival rate of the bacillus coagulans during high-temperature granulation and long-term storage, suitable for most common fermentation processes and beneficial to promoting production and application of the bacillus coagulans.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a screening method for Bacillus coagulans which is easy to form spores. Background technique [0002] Bacillus coagulans is a gram-positive bacteria that can form spores and produce lactic acid. The bacterium is rod-shaped, with blunt round ends, single or in short chains, can produce spores, and the spores are terminal. In addition to the ability of general lactic acid bacteria to maintain the balance of intestinal microecology and improve the digestive function and health level of the body, it also has the anti-stress ability of spore bacteria to resist gastric acid, choline, heat and dryness. [0003] The high stress resistance of Bacillus coagulans comes from its ability to form spores, but the spore production ability of Bacillus coagulans is far inferior to that of commonly used Bacillus such as Bacillus subtilis and Bacillus licheniformis, and its spore production ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N3/00C12N1/02C12R1/07
CPCC12N1/20C12N3/00C12N1/02
Inventor 张志榕李炯明肖俊峰帅海峰王小芬郑甜力陈志敏吴贤峰吴有林
Owner YATAI XINGYUAN FARMING TECH HAIAN CO LTD
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