A method for producing endoglucanase by fermenting roughage with mixed culture of natural symbiosis

A technology of endoglucanase and culture, which is applied in the field of biotechnology and renewable energy, can solve problems such as the inability to achieve the effect of endoglucanase with high enzyme activity, prevent bacterial contamination, facilitate promotion, The effect of improving the efficiency of anaerobic fermentation

Active Publication Date: 2021-11-02
GANSU ACAD OF SCI INST OF BIOLOGY
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Problems solved by technology

The inventor first studied the anaerobic fermentation of the co-culture of grazing yak rumen anaerobic fungi and methanobacteria using wheat straw, corn straw and rice straw as substrates (Wei Yaqin. Research on the diversity of culture and its fiber degradation characteristics [D]. 2016.), by detecting gas production, polysaccharide hydrolase activity, various esterase activities, dry matter degradation rate, phenolic acid release, methane and acetic acid production Evaluate the effectiveness of the co-culture of anaerobic fungi and methanogens in degrading straw, among which the co-cultures of anaerobic fungi and methanogens that efficiently degrade the above three straws are N.frontalis Yak16+M.ruminantium, Piromyces Yak18+M.ruminantium and O.joyonii The endoglucanase activity produced by Yak7+M.ruminantium degrading the three major straws respectively is lower than 200mU, which cannot achieve the effect of producing endoglucanase with high enzyme activity

Method used

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  • A method for producing endoglucanase by fermenting roughage with mixed culture of natural symbiosis
  • A method for producing endoglucanase by fermenting roughage with mixed culture of natural symbiosis
  • A method for producing endoglucanase by fermenting roughage with mixed culture of natural symbiosis

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Experimental program
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Effect test

Embodiment 1

[0028] Embodiment one, the preparation of mixed culture YakQH5 microbial agent

[0029] Draw 1 mL of the mixed culture YakQH5 and inoculate it into 9 mL of anaerobic medium with air-dried and pulverized wheat straw as the substrate in Heinz anaerobic tubes. ), anaerobic culture at 39°C for 72 hours, the growth peak is reached, and the fermentation broth is a high-vigor bacterial agent at this time.

Embodiment 2

[0030] Embodiment two, mixed culture YakQH5 fermentation produces endoglucanase

[0031] Put 45mL of liquid basic medium in a 100mL anaerobic fermentation bottle, and use 0.5g of dry and crushed wheat straw, corn straw, rice straw, oat straw, soybean straw, flax straw, bran, tomato residue, peanut shell and Wheat husks were used as a substrate. Deoxygenated, then autoclaved. The mixed culture YakQH5 that embodiment one obtains draws 5mL and inoculates respectively in above-mentioned anaerobic culture medium that adds substrate with sterile syringe, adds 0.5mL compound antibiotic simultaneously (1600IU / mL penicillin and 2000IU / mL streptomycin sulfate ), cultured anaerobically at 39°C for 5 days. A total of 3 parallel experiments were set up, and the enzyme activity of endoglucanase in the anaerobic bottle was measured every 24 hours.

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Abstract

The invention relates to the field of biotechnology renewable energy, in particular to a method for producing endoglucanase by fermenting roughage substrates with a natural symbiotic mixed culture YakQH5, wherein the mixed culture YakQH5 is composed of anaerobic fungi (Neocallimastix frontalis) Composed of Methanobrevibacter gottschalkii, it can degrade different roughages and produce endoglucanase with high enzymatic activity, among which the activity of endoglucanase produced by degrading wheat hulls can reach 1170mU, which has achieved remarkable results Adding compound antibiotics in the fermentation process can also prevent the mixed culture YakQH5 from being polluted by bacteria during the fermentation process, and further improve the efficiency of anaerobic fermentation. The mixed culture adopted in the present invention can be preserved in vitro for survival and passage, which is convenient for popularization and provides convenience for production.

Description

technical field [0001] The invention relates to the field of biotechnology renewable energy, in particular to a method for producing endoglucanase by fermenting roughage with natural symbiotic mixed culture. Background technique [0002] In recent years, with the increase of the population and the improvement of the quality of life in the world, the demand for energy has risen sharply. Traditional energy materials are facing the danger of depletion, and the use of these traditional energy materials produces a large amount of greenhouse gases and pollutes the environment. Alleviating the three major crises of "food, energy and environment" facing mankind today is one of the important ways to realize the sustainable development strategy of agriculture. Rough fodder such as straw, hay and husks has received more and more attention as renewable energy materials. Roughage refers to feed with a natural moisture content below 60%, crude fiber content in dry matter equal to or hig...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/42C12R1/01C12R1/645
CPCC12N9/2437C12Y302/01004
Inventor 魏亚琴毛婷王治业
Owner GANSU ACAD OF SCI INST OF BIOLOGY
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