Solid-state fermentation method for feeding bacillus subtilis with high germination rate and high stress resistance

A Bacillus subtilis, solid-state fermentation technology, applied in microorganism-based methods, biochemical equipment and methods, bacteria and other directions, can solve the problems that plague the industrial production of Bacillus subtilis, the production effect is not ideal, etc., and solve the problem of bacterial growth. Metabolic stagnation, high survival rate, effect of improving spore yield

Inactive Publication Date: 2015-03-04
ANYOU BIOTECH GRP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing culture method ferments Bacillus subtilis, and the spore rate is about 80%. After the fermentation is completed, it needs to be dried or air-dried, and the production effect is not ideal. How to increase

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] (1) Bacterial strain: Bacillus subtilis CICC 10453 was selected and purchased from China Industrial Microorganism Culture Collection Management Center;

[0029] (2) Streak the bacterial strain described in step (1) in three zones on the nutrient agar medium, and culture at 37° C. for 24 hours;

[0030] Under sterile conditions, pick a single colony and inoculate it into 20 mL of nutrient broth medium, shake it at 37°C and 180 rpm for 5 hours to obtain the primary seed liquid;

[0031] Inoculate the primary seed liquid in the nutrient broth medium with 5% inoculum amount, shake and culture at 37°C and 180rpm for 4 hours to obtain the secondary seed liquid;

[0032] The above-mentioned nutrient agar medium is composed of 0.3% beef extract, 1.0% peptone, 0.5% sodium chloride, 2.0% agar, prepared in distilled water, pH 7.2-7.4; the nutrient broth medium is composed of 0.3% beef extract, 1.0 peptone %, sodium chloride 0.5%, prepared with distilled water, pH 7.2-7.4;

[003...

Embodiment 2

[0040] (1) Bacterial strain: Bacillus subtilis CICC 10453 was selected and purchased from China Industrial Microorganism Culture Collection Management Center;

[0041] (2) Streak the bacterial strain described in step (1) in three zones on the nutrient agar medium, and culture at 37° C. for 24 hours;

[0042] Under sterile conditions, pick a single colony and inoculate it into 20 mL of nutrient broth medium, shake it at 37°C and 180 rpm for 6 hours to obtain the primary seed liquid;

[0043]Inoculate the primary seed liquid in the nutrient broth medium with 1% inoculation amount, shake and culture at 37°C and 180rpm for 6 hours to obtain the secondary seed liquid;

[0044] The above-mentioned nutrient agar medium is composed of 0.3% beef extract, 1.0% peptone, 0.5% sodium chloride, 2.0% agar, prepared in distilled water, pH 7.2-7.4; the nutrient broth medium is composed of 0.3% beef extract, 1.0 peptone %, sodium chloride 0.5%, prepared with distilled water, pH 7.2-7.4;

[0...

Embodiment 3

[0050] (1) Bacterial strain: Bacillus subtilis CICC 10453 was selected and purchased from China Industrial Microorganism Culture Collection Management Center;

[0051] (2) Streak the bacterial strain described in step (1) in three zones on the nutrient agar medium, and culture at 37° C. for 24 hours;

[0052] Under sterile conditions, pick a single colony and inoculate it into 20 mL of nutrient broth medium, shake it at 37°C and 160 rpm for 8 hours to obtain a first-grade seed liquid;

[0053] Inoculate the primary seed liquid in the nutrient broth medium with 2% inoculation amount, shake and culture at 37°C and 160 rpm for 4 hours to obtain the secondary seed liquid;

[0054] The above-mentioned nutrient agar medium is composed of 0.3% beef extract, 1.0% peptone, 0.5% sodium chloride, 2.0% agar, prepared in distilled water, pH 7.2-7.4; the nutrient broth medium is composed of 0.3% beef extract, 1.0 peptone %, sodium chloride 0.5%, prepared with distilled water, pH 7.2-7.4; ...

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Abstract

The invention discloses a solid-state fermentation method for feeding bacillus subtilis with a high germination rate and high stress resistance. The solid-state fermentation method comprises the following steps: selecting the bacillus subtilis CICC 10453; fermenting and culturing the bacillus subtilis CICC 10453 in a solid culturing medium by adopting a segmented temperature control method, wherein the main component of the solid culturing medium is wheat bran; directly collecting without the necessity of drying, and then smashing and sieving to obtain a solid finished product. Every 1g of bacillus-subtilis preparation contains 1.0*10<10>CFU effective bacillus subtilis, and more than 95% of the bacillus subtilis can germinate, so that the bacillus-subtilis preparation has good acid, cholate and high temperature resistance. If the bacillus-subtilis preparation is used as a feed additive, the unnecessary loss caused in the process of processing a feed can be reduced to a great extent. In addition, the bacillus-subtilis preparation can pass the harsh strong-acid environment in the stomach of an animal to play a beneficial role in the intestine.

Description

technical field [0001] The invention relates to the technical field of microbial fermentation engineering for feed, in particular to a solid-state fermentation method of Bacillus subtilis for feed with high spore rate and high stress resistance. Background technique [0002] Antibiotics have a high utilization rate in China, half of which are mainly used in the pharmaceutical industry, and the other half are used in the aquaculture industry. Since it was discovered in the 1950s that adding low-concentration antibiotics to feed can prevent the occurrence of animal bacterial diseases and promote animal growth, the use of antibiotics has become increasingly rampant, which has led to many food safety and biosafety problems, such as drug residues, Drug-resistant bacteria, etc. [0003] In view of a series of problems caused by the abuse of antibiotics, people gradually turned their attention to probiotics. The research on animal microecological preparations in my country starte...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12R1/125
CPCC12N1/20
Inventor 李卫娟王计伟王宇霄
Owner ANYOU BIOTECH GRP
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