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High-density mixed fermentation preparation method of composite bacillus microbial agent

A compound Bacillus and mixed fermentation technology, applied in the biological field, can solve the problems of high preparation cost, low number of effective viable bacteria, and long fermentation time, etc., to increase the number of effective viable bacteria, reduce production costs, and increase the number of spores Effect

Active Publication Date: 2020-08-25
DALIAN UNIV OF TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Aiming at the problems existing in the prior art, the purpose of the present invention is to provide a high-density mixed fermentation preparation method of a composite bacillus inoculum, aiming to solve the problems in the existing Paenibacillus polymyxa or Bacillus firmus fermentation and inoculum preparation technology. , the number of effective viable bacteria is low, the number of spores is low, the fermentation time is long, and the cost of bacterial agent preparation is high

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] The steps of applying the high-density mixed fermentation method of the present application to prepare the compound bacillus inoculum are as follows:

[0030] (1) Preparation of culture medium and dried distiller's grains enzymatic hydrolysis solution:

[0031] Activation medium: tryptone 10g / L, yeast extract 5g / L, sodium chloride 10g / L, agar 15g / L, the rest is distilled water, pH7-8.

[0032] Seed medium: tryptone 10g / L, yeast extract 5g / L, sodium chloride 10g / L, the rest is distilled water, pH7-8.

[0033] Fermentation medium: glucose 10g / L, soluble starch 10g / L, peptone 5g / L, beef extract 5g / L, yeast extract 5g / L, calcium carbonate 5g / L, and the rest is enzymatic hydrolyzate of dried distiller's grains, pH7-8 .

[0034] Feed medium: glucose 500g / L, glycerol 50g / L, and distilled water as the rest.

[0035] Dried distiller's grains enzymatic hydrolysis solution: mix the dried distiller's grains with distilled water at a mass ratio of 5%, adjust the pH to 9-11 with 1...

Embodiment 2

[0077] The steps of applying the high-density mixed fermentation method of the present application to prepare the compound bacillus inoculum are as follows:

[0078] (1) Preparation of culture medium and dried distiller's grains enzymatic hydrolysis solution:

[0079] Activation medium: tryptone 10g / L, yeast extract 5g / L, sodium chloride 10g / L, agar 15g / L, the rest is distilled water, pH7-8.

[0080] Seed medium: tryptone 10g / L, yeast extract 5g / L, sodium chloride 10g / L, the rest is distilled water, pH7-8.

[0081] Fermentation medium: glucose 20g / L, soluble starch 20g / L, peptone 15g / L, beef extract 15g / L, yeast extract 15g / L, calcium carbonate 15g / L, the rest is enzymatic hydrolyzate of dried distiller's grains, pH7-8 .

[0082] Feed medium: glucose 500g / L, glycerol 50g / L, and distilled water as the rest.

[0083] Dried distiller's grains enzymatic hydrolysis solution: mix the dried distiller's grains with distilled water at a mass ratio of 15%, adjust the pH to 9-11 with ...

Embodiment 3

[0125] The steps of applying the high-density mixed fermentation method of the present application to prepare the compound bacillus inoculum are as follows:

[0126] (1) Preparation of culture medium and dried distiller's grains enzymatic hydrolysis solution:

[0127] Activation medium: tryptone 10g / L, yeast extract 5g / L, sodium chloride 10g / L, agar 15g / L, the rest is distilled water, pH7-8.

[0128] Seed medium: tryptone 10g / L, yeast extract 5g / L, sodium chloride 10g / L, the rest is distilled water, pH7-8.

[0129] Fermentation medium: glucose 15g / L, soluble starch 15g / L, peptone 10g / L, beef extract 10g / L, yeast extract 10g / L, calcium carbonate 10g / L, the rest is enzymatic hydrolyzate of dried distiller's grains, pH7-8 .

[0130] Feed medium: glucose 500g / L, glycerol 50g / L, and distilled water as the rest.

[0131] Dried distiller's grains enzymatic hydrolysis solution: mix the dried distiller's grains with distilled water at a mass ratio of 10%, adjust the pH to 9-11 with ...

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Abstract

The invention belongs to the technical field of biology, and particularly relates to a high-density mixed fermentation preparation method of composite bacillus microbial agent. Rich nutrient components of the dried distillers' grains are utilized, enzymatic hydrolysate generated by alkaline protease enzymolysis of the dried distillers' grains serves as a fermentation culture medium solvent, and high-density mixed fermentation of the biocontrol probiotic paenibacillus polymyxa and the bacillus firmus is achieved by controlling the fermentation pH, performing fed-batch cultivation and maintaining the dissolved oxygen amount. The effective viable count of the paenibacillus polymyxa in the fermentation liquor can reach 1.2 to 1.5* 10<10>CFU / mL, and the effective viable count of the bacillus firmus can reach 8.0 to 9.0*10<9> CFU / mL, so that the fermentation time is shortened, and the production efficiency is improved. The number of effective viable bacteria or viable spores in the compositebacillus microbial agent prepared from the dry vinasse enzymolysis residues can reach 3.0-6.6*10<11>CFU / g, full utilization of the dry vinasse raw material is achieved, and the production cost of themicrobial agent is reduced.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to a high-density mixed fermentation preparation method of a compound bacillus bacterial agent. Background technique [0002] Paenibacillus polymyxa and Bacillus firmus are Gram-positive bacteria of the genus Bacillus, which are easy to cultivate in aerobic or facultative anaerobic environments, can produce spores, and are harmless to humans, animals and plants. Pathogenicity, has been listed as an important biological pesticide and biological control agent by many countries or regions. Among them, Paenibacillus polymyxa is listed as a first-class strain exempted from safety identification by my country, and it is also one of the commercially applied microorganisms approved by the U.S. Environmental Protection Agency, while Bacillus firmus has been approved by the Brazilian National Health Supervision Bureau as a biocidal agent. nematode or seed treatment. Paenibaci...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N3/00C12R1/12C12R1/07
CPCC12N1/20C12N3/00
Inventor 薛闯吴又多李苗苗
Owner DALIAN UNIV OF TECH
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