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Strain domestication method for improving bacillus spore production rate

A technology for bacillus and spore production, which is applied in the field of microorganisms, can solve the problems of long culture time and complicated operation of bacillus, and achieve the effects of shortening the cultivation and domestication time, simple operation and increasing the spore production rate

Active Publication Date: 2018-05-04
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved in the present invention is to provide a method for acclimating strains to improve the spore-producing rate of bacillus in order to overcome the problems in the prior art of breeding bacillus with high spore-yielding rate, long cultivation time and complicated operation

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] 1. Preparation of seed solution

[0031] Ferment the preserved Bacillus subtilis ANSB060 shake flask to make seed liquid, and ferment and cultivate the shake flask in a 37°C incubator for 12 hours at a rotation speed of 200r / min to obtain Bacillus subtilis ANSB060 seed liquid (the concentration of viable bacteria is about 10 9 CFU / mL)

[0032] The formula of the fermentation medium is: every 1000mL medium contains: tryptone 8g, yeast extract 3g, glucose 2g, beef extract 5g, sodium chloride 3g, disodium hydrogen phosphate 4g, magnesium sulfate heptahydrate 1.5g, pH is 7.0.

[0033] After the above-mentioned ingredients are weighed according to the formula, dissolve in an appropriate amount of water, then add water to adjust the pH value, and sterilize by steam at 121°C for 20 minutes, and set aside.

[0034] 2. Subculture of strains with different concentrations of medium

[0035] Get above-mentioned Bacillus subtilis ANSB060 seed solution and inoculate in the 250ml E...

Embodiment 2

[0045] 1. Preparation of seed solution

[0046] The preserved Bacillus subtilis ANSB01G was fermented in a shake flask to make a seed solution, and cultured in a 37° C. incubator with shaking for 12 hours at a rotation speed of 200 r / min to obtain a Bacillus subtilis ANSB01G seed solution. (The concentration of live bacteria is about 10 9 CFU / mL)

[0047] The formula of the fermentation medium is: every 1000mL medium contains: 10g of tryptone, 2g of yeast extract, 2g of glucose, 3g of beef extract, 4g of sodium chloride, 3g of disodium hydrogen phosphate, 1.0g of magnesium sulfate heptahydrate, pH The value is 7.2. The preparation method is as in Example 1, steam sterilization at 121° C. for 20 minutes.

[0048] 2. Subculture of strains with different concentrations of medium

[0049] Get the above-mentioned bacillus subtilis ANSB01G seed liquid and inoculate in the 250ml Erlenmeyer flask of 50ml original concentration fermented liquid (the fermented medium of above-mentio...

Embodiment 3

[0060] 1. Preparation of seed solution

[0061] The preserved Bacillus subtilis ANSB471 was fermented in a shaker flask to make a seed solution, and cultured in a 37°C incubator with shaking for 12 hours at a speed of 200r / min to obtain a Bacillus subtilis ANSB471 seed solution (the concentration of viable bacteria was about 10 9 CFU / mL).

[0062] The formula of the fermentation medium is: every 1000mL medium contains: tryptone 12g, yeast extract 1.5g, glucose 3g, beef extract 2g, sodium chloride 4g, disodium hydrogen phosphate 2.5g, magnesium sulfate heptahydrate 0.5g, Distilled water 1000ml, pH value 7.4; preparation method as in Example 1, steam sterilization at 121°C for 20min.

[0063] 2. Subculture of strains with different concentrations of medium

[0064] Get the above-mentioned Bacillus subtilis ANSB471 inoculated in the 250ml Erlenmeyer flask of 50ml original concentration fermented liquid (((the fermented medium of above-mentioned " 1, in the preparation of seed l...

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Abstract

The invention discloses a strain domestication method for improving the bacillus spore production rate. The method comprises the following steps of activating bacillus; carrying out sub-culturing on the activated bacillus according to a sequence from a normal culture medium to a poor culture medium, and then increasing nutritive material concentration to culturing in the normal culture medium sequentially, wherein three culture media or more are set according to the concentration, and continuous passage is carried out for three generations or more. According to the strain domestication methodfor improving the bacillus spore production rate provided by the invention, the culture and domestication time is shortened to a great extent, the method is simple to operate, and the bacillus spore production rate can be improved to 96 percent according to the method provided by the invention.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a strain domestication method for improving the spore-producing rate of bacillus. Background technique [0002] As microecological preparations have become a hot spot in the research of feed additives, the advantages of probiotic preparations in animal feeding have gradually emerged. However, most probiotics have the disadvantages of poor stress resistance and heat resistance, which bring great difficulties to the preservation and transportation of preparations. Bacillus is an aerobic spore-forming bacterium that widely exists in nature. Bacillus is a dormant body with extremely low water content and strong stress resistance formed in the cell when the necessary nutrients of bacteria are about to be exhausted. [0003] Studies have shown that the spores produced by Bacillus have various stress resistances such as heat resistance, drought resistance, UV resistance and orga...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/36C12R1/125
CPCC12N1/36
Inventor 马秋刚赵丽红计成李美玲
Owner CHINA AGRI UNIV
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