Trichoderma atroviride strain and application thereof in preparation of cellulase

A technology of Trichoderma viridans and cellulase, applied in the direction of biochemical equipment and methods, enzymes, enzymes, etc., to achieve the effects of high activity, short fermentation cycle, and reasonable components

Active Publication Date: 2013-02-13
郑州运维生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is still a certain distance from industrial application

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  • Trichoderma atroviride strain and application thereof in preparation of cellulase

Examples

Experimental program
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Effect test

Embodiment 1

[0024] Example 1 Trichoderma dark green B-8-1-34 liquid fermentation enzyme production

[0025] After the activation of the Trichoderma dark green B-8-1-34 bacterial strain whose strain preservation number is CGMCC No. 6390, 10 6 mL of spore suspension, inoculate 5% of the inoculum into the seed medium, culture at 30°C, 180rpm shaker for 24 hours, then inoculate the liquid enzyme production medium with 10% inoculum (filling volume 40 mL / 250mL), with an initial pH of 4.0-6.0, cultured at 30°C, 200rpm shaker to 66, and centrifuged at 4000r / min for 10min to prepare crude enzyme solution.

[0026] The composition of the seed medium: carboxymethyl cellulose 1%, peptone 0.2%, (NH 4 ) 2 SO 4 0.3%, MgSO 4 0.04%, CaCl 2 0.04%, KH 2 PO 4 0.1%, the balance is distilled water. Each component is expressed in weight percent.

[0027] The composition of the liquid fermentation medium: 2.5% of corn stalks, corn cobs, wheat straw or xylose residue and 0.5% of rice straw, and adde...

Embodiment 2

[0032] Embodiment 2 Trichoderma dark green B-8-1-34 solid state fermentation produces enzyme

[0033] The concentration made by activating the Trichoderma dark green strain with the strain preservation number of CGMCC No. 6390 is 10 8 The spore suspension per ml was inserted into the seed medium, cultivated in a shaker at 30°C and 180rpm for 24h, and then connected to the solid fermentation medium with 3% inoculum for cultivation, and the material was turned over regularly. 10 times the volume of 0.05 mol / L citric acid-sodium citrate buffer solution with pH 5.0, shaking and extracting at 150 rpm, and centrifuging at 4000 r / min for 10 min to prepare the crude enzyme solution.

[0034] The composition of the seed medium: carboxymethyl cellulose 1%, peptone 0.2%, (NH 4 ) 2 SO 4 0.3%, MgSO 4 0.04%, CaCl 2 0.04%, KH 2 PO 4 0.1%, the balance is distilled water. Each component is expressed in weight percent.

[0035] The composition of the solid fermentation medium: co...

Embodiment 3

[0040] Example 3 Application of saccharified lignocellulose

[0041] In 0.05mol / L acetic acid-sodium acetate or citric acid-sodium citrate buffer solution with pH 5.0, add the crude enzyme liquid or enzyme powder prepared above to corncobs, rice straw powder or xylose residue, and the ratio of solid to liquid is 1:10, 50°C, 180rpm hydrolysis for 48 hours, then measure reducing sugar concentration and glucose concentration, the results are shown in Table 3. . The amount of enzyme added is 4FPA / ml. Lignocellulose and enzymes can be added all at once or in batches.

[0042] Table 3 Results of saccharified lignocellulose

[0043]

[0044] Note: The DNS method is used for the determination of reducing sugar; the enzyme method is used for the determination of glucose (SBA-40c biosensor analyzer)

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Abstract

The invention relates to a trichoderma atroviride B-8-1-34 strain capable of highly yielding cellulose and application of the trichoderma atroviride B-8-1-34 strain in the preparation of the cellulose, belonging to the technical field of microorganisms. The trichoderma atroviride B-8-1-34 strain disclosed by the invention has been preserved in China General Microbiological Culture Collection Center on Aug.10, 2012, with the preservation number of CGMCC No.6390. The trichoderma atroviride B-8-1-34 disclosed by the invention can be used for producing the cellulose by making use of lignocellulose-containing industrial and agricultural wastes in combination of low-cost raw materials such as bran, maize slurry and soybean cake powder through liquid or solid fermentation, has the advantages of short fermentation period, high activity of produced cellulose, reasonable components and high lignocellulose hydrolysis, and provides a novel strain resource to solve the problems on high production cost, low saccharifying efficiency and the like caused by low enzyme activity and unreasonable enzyme components in the utilization of cellulose resources.

Description

technical field [0001] The invention relates to a Trichoderma dark green B-8-1-34 strain with high cellulase production and its application in preparing cellulase, belonging to the technical field of microorganisms. Background technique [0002] Cellulose is one of the most abundant renewable resources in nature. Degrading natural cellulose into usable sugars, and further converting them into ethanol, bacterial protein, gas fuel and other substances, has great practical significance for solving the problems of environmental pollution, food shortage, resource shortage and energy crisis faced by the world today. Significance, and cellulase is the most effective biocatalyst for degrading cellulose. [0003] The high cost of enzymatic hydrolysis is one of the main constraints to the industrialization of biomass energy. Reducing the cost of cellulase mainly involves reducing the culture medium and fermentation cost of cellulase-producing bacteria, and improving the activity of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N9/42C12P19/14C12R1/885
Inventor 戴桂馥吴健刘允攀师会勤高明夫房晓敏潘静
Owner 郑州运维生物技术有限公司
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