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Bacillus licheniformis for degrading cellulose and application thereof in straw fermentation

A technology of Bacillus licheniformis and plant cellulose, which is applied in the direction of application, bacteria, and microbial-based methods, can solve problems such as less research, and achieve the effects of increasing utilization, improving the internal environment of animals, and good thermal stability

Inactive Publication Date: 2008-07-02
THE INST OF MICROBIOLOGY XINJIANG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, most Bacillus licheniformis at home and abroad are used to produce protease and amylase, but Bacillus licheniformis has a special effect on cellulose decomposition. expression, but there are few studies on its production application, especially in the production of silage and yellow silage. It is about the production of cellulase and stable growth under low pH conditions, which can be directly applied to silage and yellow silage For feed, it has not been reported that it has a good degradation effect on plant fiber

Method used

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  • Bacillus licheniformis for degrading cellulose and application thereof in straw fermentation
  • Bacillus licheniformis for degrading cellulose and application thereof in straw fermentation
  • Bacillus licheniformis for degrading cellulose and application thereof in straw fermentation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Embodiment 1: the separation of bacterial classification

[0045] 1: Take a certain amount of sample and wash it with sterile water, and draw a certain amount of suspension and spread it on the LB plate containing carboxymethyl cellulose.

[0046] 2: Separate a single colony after two to three days, then stain the plate with Congo red (1g / L) for about one hour, and then decolorize it with sodium chloride (1mol / L).

[0047] 3: After decolorization, observe whether there is a transparent circle, and then preliminarily screen out six strains with the ability to decompose cellulose through the ratio of the colony size to the transparent circle.

[0048] 4: Preliminary fermentation enzyme production research, through the supernatant of the fermentation broth and the supernatant after breaking the wall, 8000rpm, 5min, take the supernatant, and 3,5-dinitrosalicylic acid color development (DNS method) to preliminarily determine the enzyme-producing genus Inner or extracellular...

Embodiment 2

[0051] Example 2: Growth characteristics of bacterial strain B.licheni ws6

[0052] 1: Cultivate with LB liquid medium, first set up four temperature gradients, respectively 30°C, 40°C, 50°C, 60°C, according to the inoculation amount of 1%, 100rpm, sampling once every two hours, measured under the condition of λ540nm OD value. Conclusion It can grow at 30-50℃. It stops growing at 60°C and grows rapidly between 30°C and 40°C. Furthermore, three gradients were set up, respectively 30°C, 35°C, and 40°C, and the optimum growth temperature was determined to be 35-40°C.

[0053] 2: Observation by puncture culture of LB solid medium, growth everywhere in the puncture tube, the strain belongs to facultative anaerobic bacteria.

[0054] 3: Cultivate with LB liquid medium, set the medium at pH 4, 5, 6, 7, 8, use 1% inoculum, 100 rpm, take samples every two hours, measure OD value under the condition of λ540nm, conclusion: It grows well around pH 4.0 to 5.0, and also grows at pH 8.0....

Embodiment 3

[0055] Example 3: Enzyme Activity Characteristics of Bacterial Strain B.licheni ws-6

[0056] 1: Inoculate at 1% inoculum size, use LB liquid medium containing carboxymethyl cellulose as the primary fermentation medium, 100rpm, 37°C, sample once every two hours, 8000rpm, 5min, take the supernatant with carboxymethylcellulose Cellulose is used as the substrate, and the optimum time for enzyme production is measured to be 36 to 38 hours by DNS chromogenic method.

[0057] 2: Set up four gradients, respectively 30°C, 40°C, 50°C, and 60°C. The optimum temperature is 40°C (the method is the same as above), the time is 20min, and the enzyme activity is about 1300mu / ml.

[0058] 3: Set up four gradients, the pH is 3, 4, 5, 6, 7, and the optimum substrate pH is measured between 4-5 (the method is the same as above, and the substrate concentration is 0.1g / ml.

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Abstract

The invention discloses a Bacillus licheniformis ws-6 capable of effectively decomposing plant fiber, which is a probiotics belonging to Bacillus licheniformis by identification. The bacterium can be directly used for production of feedstuff additives and can effectively decompose cellulose in green and yellow sillage, which facilitates absorption and transformation by animals, thus effectively improving the utilization rate of the feedstuff and significantly improving the internal environment of animals. The Bacillus licheniformis ws-6 belongs to microbe with four-stage safety for human and animals, and can be widely used in the field of fermentation of straw.

Description

field of invention [0001] The invention relates to the field of microbial fermentation and its product enzymes. Specifically, the invention relates to a cellulose-degrading bacillus licheniformis (Bacillus licheniformis) and its application in straw fermentation. Background technique [0002] Cellulase refers to the general term for a class of enzymes that can degrade cellulose. It is a complex enzyme system composed of a variety of hydrolytic enzymes, mainly from fungi and bacteria. Most of the cellulose-producing bacteria used in current research and production are Trichoderma, Aspergillus and Penicillium, etc., because fungi that can decompose crystalline cellulose can secrete more or less cellulase, so the fungal source of cellulase Still relatively broad. Some bacteria can also decompose cellulose and produce cellulase, but because the amount of cellulase secreted is low, and most of them are intracellular enzymes or adhere to the cell wall, it is difficult to carry o...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12S3/04A23K1/14C12R1/10
Inventor 王炜詹发强包慧芳崔春生崔卫东龙宣杞
Owner THE INST OF MICROBIOLOGY XINJIANG ACADEMY OF AGRI SCI
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