Preparation method for lactic acid-producing bacillus subtilis microbial preparation

A technology of Bacillus subtilis and microbial preparations, applied in the field of microorganisms, can solve the problems of high rate of miscellaneous bacteria and large fluctuation of the number of viable bacteria, and achieve the effect of increasing the number of viable bacteria

Inactive Publication Date: 2012-02-01
GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the preparation of Bacillus subtilis microbial preparations generally adopts the traditional fermentation process of open bran solid state, using natural wind, natural temperature and humidity. In the prepared Bacillus subtilis microbial preparations, the rate of miscellaneous bacteria is high, and the number of viable bacteria fluctuates greatly.

Method used

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  • Preparation method for lactic acid-producing bacillus subtilis microbial preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Pick a ring of bacteria from the freeze-dried seeds of Bacillus subtilis and inoculate it into the slant medium of beef extract peptone agar test tube, and incubate at 37°C for 16 hours; then transfer the slant seeds of the test tube to the slant medium of beef extract peptone agar eggplant bottle In the medium, cultured at 37°C for 24 hours, after testing, more than 80% of them formed spores. Scrape and wash the mature fungus mud on the eggplant bottle with sterile water, put it into a 250ml triangular flask, oscillate to make the bacteria uniform, and form a seed culture solution with a concentration of 1.2×10 9 cfu / ml. The formula of the beef extract peptone agar medium is as follows: each liter contains 10 g of peptone, 5 g of beef extract, 5 g of NaCl, 15 g to 20 g of agar, the balance is water, pH 7.2 to 7.4, and autoclaved at 121° C. for 20 minutes.

[0025] Transfer the seed culture solution to a 30L seed tank with a primary fermentation medium with a 5% inocul...

Embodiment 2

[0055] Pick a ring of bacteria from the freeze-dried seeds of Bacillus subtilis and inoculate it into the slant medium of beef extract peptone agar test tube, and incubate at 37°C for 12 hours; then transfer the slant seeds of the test tube to the slant medium of beef extract peptone agar eggplant bottle In culture, cultured at 37°C for 30 hours, after testing, more than 80% of the bacteria formed spores. Scrape and wash the mature fungus mud on the eggplant bottle with sterile water, put it into a 250ml triangular flask, oscillate to make the bacteria uniform, and form a seed culture solution with a concentration of 2.0×10 9 cfu / ml. The formula of the beef extract peptone agar medium is as follows: each liter contains 10 g of peptone, 5 g of beef extract, 5 g of NaCl, 15 g to 20 g of agar, the balance is water, pH 7.2 to 7.4, and autoclaved at 121° C. for 20 minutes.

[0056] Transfer the seed culture solution to a 30L seed tank with a primary fermentation medium with a 3% i...

Embodiment 3

[0062] Pick a ring of bacteria from the freeze-dried seeds of Bacillus subtilis and inoculate it into the slant medium of beef extract peptone agar test tube, and incubate at 37°C for 16 hours; then transfer the slant seeds of the test tube to the slant medium of beef extract peptone agar eggplant bottle In the medium, cultured at 37°C for 24 hours, after testing, more than 80% of them formed spores. Scrape and wash the mature fungus mud on the eggplant bottle with sterile water, put it into a 250ml triangular flask, oscillate to make the bacteria uniform, and form a seed culture solution, the concentration of the bacteria in the seed solution is 5.0×10 8 cfu / ml. The formula of the beef extract peptone agar medium is as follows: each liter contains 10 g of peptone, 5 g of beef extract, 5 g of NaCl, 15 g to 20 g of agar, the balance is water, pH 7.2 to 7.4, and autoclaved at 121° C. for 20 minutes.

[0063] Transfer the seed culture solution to a 30L seed tank with a primary f...

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Abstract

The invention discloses a preparation method for lactic acid-producing bacillus subtilis microbial preparation. After undergoing primary fermentation and secondary fermentation, a seed culture solution is then transferred into a bran solid medium, and is fermented under appropriate conditions, and thereby the lactic acid-producing bacillus subtilis microbial preparation is prepared. The preparation method optimizes a fermentation medium, a thallus as the seed culture solution, more than 80 percent of which are formed into spores, is inoculated into the fermentation medium, and is subjected tothe primary fermentation and the secondary fermentation under the appropriate conditions, a secondary seed solution is then inoculated into the bran solid medium, and is controlled to ferment at highdensity under appropriate conditions, so that a large quantity of spores can be formed, and meanwhile, the affection of foreign bacteria is prevented. The preparation method effectively solves the problems of the bran fermentation process in the prior art, such as excessive foreign bacteria, uneven ventilation and heat dissipation, non-constant temperature and humidity and the like, and greatly increases the viable count of the bacilli subtilis; in a test, the viable count per gram of the lactic acid-producing bacillus subtilis microbial preparation prepared by the method is 20 to 25 billion,the spore rate is larger than 80 percent, and the foreign bacteria rate is 0.5 to 2 percent.

Description

Technical field: [0001] The invention belongs to the field of microorganisms, and in particular relates to a preparation method of a lactic acid-producing Bacillus subtilis microbial preparation. Background technique: [0002] Bacillus subtilis (Bacillus subtilis) is one of the bacillus allowed to be used as a feed additive by the Ministry of Agriculture of my country, and it has been increasingly developed into feed microbial preparations. Bacillus subtilis has strong protease, lipase, amylase and other activities, can produce antibiotics, has strong biological oxygen capture ability in animal intestines, and can produce L-lactic acid under anaerobic conditions. These characteristics play an important role in promoting the digestion and absorption of animal nutrition, improving the feed conversion rate of animals, preventing diseases and promoting growth. [0003] Therefore, adding Bacillus subtilis capable of producing L-lactic acid in feed is a very green and healthy cho...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A23K1/16C12N1/20C12R1/125A23K10/18
Inventor 许国焕江永明吴月嫦张丽陈亚剑郭莹姿郭慧青王珺董小林刘海龙
Owner GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY
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