Method for producing viable bacteria of bacillus licheniformis by high-density solid fermentation

A technology of bacillus licheniformis and solid fermentation, applied in the biological field, can solve the problems of difficult control of moisture and temperature, difficult control of miscellaneous bacteria pollution, difficult control of fermentation process, etc., to meet the nutritional needs of bacteria breeding and increase the content of effective live bacteria , low cost effect

Active Publication Date: 2014-07-30
DONGGUAN BAODE BIOLOGICAL ENG
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The production of Bacillus licheniformis mainly includes liquid fermentation and solid fermentation. Liquid fermentation requires a complete set of fermenter equipment. Its advantages are that the fermentation process is easy to control and the product has high purity. The disadvantage is that it needs to be adsorbed and dried after fermentation. High energy consumption and serious environmental pollution; solid fermentation is generally carried out in the fermentation room. Bacterial contamination is difficult to control
[0005] The Chinese patent application number is 201110324147.2, which discloses a method for producing live bacteria of Bacillus subtilis by solid fermentation, but the strains used, the specific method of seed cultivation, the specific operation steps of solid fermentation cultivation, and the fermentation products are air-dried. The drying method and the formula of the medium used are different, and the operation method of the reference document is complicated and the production efficiency is low
Due to the heat generated during the fermentation process, the temperature of the material rises rapidly, and the water and temperature are not easy to control in the general open culture method.

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0037] A kind of high-density solid fermentation method of the present embodiment produces bacillus licheniformis, comprises the following steps:

[0038] (1) Selection of strains: Bacillus licheniformis ( Baclicus lincheniformis ) CGMCC1.0518 strain;

[0039] (2) Slant seed culture: inoculate the strain described in step (1) on the slant surface of nutrient agar medium under sterile conditions, and cultivate at 30°C for 48 hours to obtain slant seed;

[0040] (3) Cultivation of solid seeds: Wash the slanted seeds with sterile water, collect them in a triangular flask, and count them by serial dilution. The viable bacteria content of the bacterial suspension is 2×10. 9 / mL, diluted with sterile water to 1 × 10 8 / mL, and then with the inoculum of 5% v / w (volume-to-mass ratio), the bacterial solution was inoculated into the solid medium sterilized at 121 ° C for 90 min (1L solid bacteria bottle was filled with 180 g of wet material), 30 ° C Cultivated for 60 hours to obtain so...

Embodiment 2

[0047] A method for producing Bacillus licheniformis by high-density solid fermentation of the present embodiment comprises the following steps:

[0048] (1) Selection of strains: Bacillus licheniformis ( Baclicus lincheniformis ) CGMCC1.0518 strain;

[0049] (2) Slant seed culture: inoculate the strain described in step (1) on the slant surface of nutrient agar medium under sterile conditions, and cultivate at 30°C for 48 hours to obtain slant seed;

[0050] (3) Cultivation of solid seeds: Wash the slanted seeds with sterile water, collect them in a triangular flask, and count them by serial dilution. The viable bacteria content of the bacterial suspension is 1.8×10. 9 / mL, diluted with sterile water to 1 × 10 8 / mL, and then with the inoculum volume of 5% v / w (volume to mass ratio), the bacterial solution was inoculated into the solid medium sterilized at 121 ° C for 100 min (1L solid bacteria bottle was filled with 190 g of wet material), 32 ° C Cultivated for 55 hours t...

Embodiment 3

[0057] A method for producing Bacillus licheniformis by high-density solid fermentation of the present embodiment comprises the following steps:

[0058] (1) Selection of strains: Bacillus licheniformis ( Baclicus lincheniformis ) CGMCC1.0518 strain;

[0059] (2) Slant seed culture: inoculate the strain described in step (1) on the slant surface of nutrient agar medium under sterile conditions, and cultivate at 30°C for 48 hours to obtain slant seed;

[0060] (3) Cultivation of solid seeds: Wash the slanted seeds with sterile water, collect them in a triangular flask, and count them by serial dilution. The viable bacteria content of the bacterial suspension is 2×10. 9 / mL, diluted with sterile water to 1 × 10 8 / mL, and then with the inoculation amount of 5% v / w (volume-to-mass ratio), the bacterial solution was inoculated into the solid medium sterilized at 121 ° C for 120 min (200 g of wet material in 1 L solid bacteria bottle), 40 ° C Cultivated for 48 hours to obtain so...

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Abstract

The invention relates to the field of biotechnology, in particular to a method for producing viable bacillus licheniformis by high-density solid fermentation, which comprises the following steps: strain selection, slant seed cultivation, solid seed cultivation, solid fermentation cultivation, fermentation product drying and The fermentation product is flash evaporated. The raw materials of the culture medium of the present invention are all agricultural by-products, the cost is low, the formula is reasonable, and the nutritional requirements for the propagation of bacteria can be met. Especially adding the pH value change of medium in the calcium carbonate buffer culture process can greatly improve the effective viable bacteria content of bacillus licheniformis of solid fermentation, and experiment and production prove that the effective viable bacteria number of bacillus licheniformis prepared by the inventive method can reach Up to 100 billion / g, the rate of miscellaneous bacteria in the product is less than 5%, and the rate of spores is high, reaching more than 90%, which can greatly reduce the production cost of enterprises and meet the needs of microbial bacterial agent production.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for producing live bacillus licheniformis by high-density solid fermentation. Background technique [0002] Bacillus licheniformis ( Baclicus lincheniformis ) is a type of Bacillus, single cell, 0.6~0.8×1.5~3 microns, cell shape and arrangement are rod-shaped, solitary, no poly-β-hydroxybutyrate particles in the cell, Gram-positive bacillus . Produce mesophilic ellipsoid spores with slightly enlarged cysts. The colonies are flat, irregularly bordered, and white. [0003] Bacillus licheniformis ( Baclicus lincheniformis ) has multiple functions and is an excellent strain that can be used in many fields, such as microbial pesticides, microbial feed additives, microbial fertilizers, water purifiers, etc. Its main function is to inhibit the growth of harmful bacteria (Escherichia coli, Clostridium perfringens), increase the beneficial bacteria in the body and reduce the pot...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12R1/10
Inventor 张新雄彭锋毛光平张东升常留群
Owner DONGGUAN BAODE BIOLOGICAL ENG
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