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Method for producing viable bacillus subtilis by way of solid fermentation

A Bacillus subtilis, solid fermentation technology, applied in the biological field, can solve the problems of many process procedures, difficult control of bacterial pollution, difficult control of fermentation process, etc., and achieves the effects of low environmental pollution, increased effective viable bacteria content and low cost.

Inactive Publication Date: 2012-02-15
山东省农业科学院高新技术研究中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The production of Bacillus subtilis 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 adsorption and drying after fermentation, and there are many process procedures; It is carried out in the fermentation room. The advantage is that the fermentation yield is high and no adsorption is required after fermentation. It can be used directly after drying. The process is simple. The disadvantage is that the fermentation process is not easy to control, and the contamination of bacteria is not easy to control.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] (1) Strain selection: select Bacillus subtilis (Bacillus subtilis) ACCC03120 bacterial strain; Described strain is purchased from China Agricultural Microorganism Strain Collection and Management Center, has nitrogen fixation and crop rhizosphere growth-promoting ability;

[0021] (2) Seed culture: put the strain described in step (1) into a 100 mL liquid seed medium with an inoculation loop under aseptic conditions, and culture with shaking at 37° C. for 28 hours to obtain a first-grade seed liquid;

[0022] (3) Shake flask fermentation culture:

[0023] With the inoculum amount of 5% volume ratio, inoculate the primary seed liquid in the shake flask that 1000mL liquid seed culture medium is housed, 37 ℃, shake the flask fermentation with the rotating speed of 180 revs / min for 28 hours, the fermentation finishes, obtains two grade seed liquid;

[0024] (4) Bacterial colony counting: Carry out plate counting with the serial dilution method, the live bacteria content of...

Embodiment 2

[0027] (1) strain selection: select Bacillus subtilis (Bacillus subtilis) ACCC03120, and the strain is purchased from China Agricultural Microorganism Culture Collection Management Center;

[0028] (2) Seed culture: the bacterial strain described in step (1) was placed in 60 mL of liquid fermentation seed medium with an inoculation loop under aseptic conditions with 1 to 2 loops, and cultured with shaking at 37° C. for 30 hours to obtain a first-grade seed liquid;

[0029] (3) Shake flask fermentation culture:

[0030] With the inoculum amount of 3% volume ratio, the primary seed liquid is inoculated in the shake flask that 1000mL liquid seed culture medium is housed, 37 ℃, with the rotating speed shake bottle fermentation of 180 rev / mins 28 hours, fermentation finishes, obtains two grade seed liquid;

[0031] (4) Solid fermentation culture:

[0032] With the inoculum amount of 10% mass ratio, inoculate the secondary seed solution in 0.2 tons of solid medium that has been m...

Embodiment 3

[0039] (1) Strain selection: select Bacillus subtilis ACCC03120;

[0040] (2) Seed culture: put the strain described in step (1) into 100 mL liquid seed culture medium with an inoculation loop under aseptic conditions, and culture with shaking at 37° C. for 30 hours to obtain a first-grade seed liquid;

[0041] (3) Shake flask fermentation culture:

[0042] With the inoculum amount of 2% volume ratio, the primary seed liquid is inoculated in the shake flask that 1000mL liquid seed culture medium is housed, 37 ℃, with the rotating speed shake bottle fermentation of 180 rev / mins 28 hours, fermentation finishes, obtains two grade seed liquid;

[0043] (4) Solid fermentation culture:

[0044] With an inoculation amount of 7% by mass, inoculate the secondary seed solution in 0.8 tons of solid medium that has been mixed and sterilized in advance, and spread the inoculated solid medium on the bottom belt with a thickness of 7 cm. In the plastic baskets in the gap, place the plasti...

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Abstract

The invention discloses a method for producing viable bacillus subtilis by way of solid fermentation, which includes the following steps: the ACCC03120 bacillus subtilis strain is chosen, cultured by utilizing solid fermentation under the temperature of 30 DEG C to 35 DEG C for 48 to 72 hours, then naturally aired for 35 to 45 hours and ground to pass a 80-mesh sieve, and thereby a finished solidfermentation product containing the viable bacillus subtilis is prepared. The material cost of the method is low, the production process is simple, large equipment is not needed, moreover, the effective viable count of the bacillus subtilis in the product is high and 60 billion per gram of the dry fermentation product, and the demand on a great deal of microorganism fertilizer can be satisfied.

Description

technical field [0001] The invention relates to a method for producing live bacillus subtilis (Bacillus subtilis) bacteria by solid fermentation, which belongs to the field of biotechnology. Background technique [0002] Bacillus subtilis is a kind of Bacillus, single cell, 0.7~0.8×2~3 micron, non-capsulated, perinatal flagella, able to move, Gram-positive bacteria, spore 0.6~0.9×1.0 ~1.5 microns, oval to columnar, located in the center of the fungus or slightly off, the fungus does not expand after spore formation. The surface of the colony is rough and opaque, stained white or yellowish, and often forms wrinkles when growing in liquid medium. [0003] Bacillus subtilis (Bacillus subtilis) has multiple functions and is an excellent strain that can be applied in many fields, such as microbial pesticides, microbial feed additives, bio-organic fertilizer inoculants, water purifiers, etc. Its main functions are to inhibit the growth of harmful bacteria (Escherichia coli, Salm...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/125
Inventor 游银伟岳寿松尤升波柳絮范仲学张斌
Owner 山东省农业科学院高新技术研究中心
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