Trichoderma reesei strain capable of producing cellulase with high yield through space mutation

A technology of Trichoderma reesei and cellulase, applied in the direction of enzymes, enzymes, fungi, etc., can solve the problems of high blindness, heavy workload, and many test materials, and achieve the effect of broad industrial application prospects

Inactive Publication Date: 2015-02-04
河南天冠纤维乙醇有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As we all know, the method of using ultraviolet mutagenesis is a common method for strain selection, but it has certain shortcomings, such as: less beneficial variation, heavy workload, high blindness, more test materials, etc.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] 1) Preliminary screening: The spore freeze-dried powder after space mutagenesis was gradiently diluted with sterile saline to make the spore concentration 10 3 cells / mL, take 0.05mL of the diluted sample, spread it on sterile cellulose-Congo red screening medium (121°C, sterilize for 30min), culture at 28°C for 36h, and select the transparent circle / colonies with larger diameter 1000 bacterial strains; wherein, the composition of the cellulose-Congo red screening medium is uniform: microcrystalline cellulose 10g, glucose 2g, agar 18g, Congo red 0.2g, Mandels inorganic nutrient salt 1000mL, pH value natural;

[0029]2) Re-screening: The 1,000 strains selected in step 1) were inserted into the PDA slant medium, and the spores were covered with the slant, and the spores were washed with sterile saline to prepare 10 strains. 7 Each / mL spore suspension was inserted into the enzyme production medium (50mL / 250mL liquid volume) of the Erlenmeyer flask with 10% inoculum volume (...

Embodiment 2

[0039] 1) Preliminary screening: The spore freeze-dried powder after space mutagenesis was gradiently diluted with sterile saline to make the spore concentration 10 3 cells / mL, take 0.05mL of the diluted sample, spread it on sterile cellulose-Congo red screening medium (121°C, sterilize for 30min), culture at 30°C for 42h, and pick out the transparent circle / colonies with larger diameter 1000 bacterial strains; wherein, the composition of the cellulose-Congo red screening medium is uniform: microcrystalline cellulose 10g, glucose 2g, agar 18g, Congo red 0.2g, Mandels inorganic nutrient salt 1000mL, pH value natural;

[0040] 2) Re-screening: The 1,000 strains selected in step 1) were inserted into the PDA slant medium, and the spores were covered with the slant, and the spores were washed with sterile saline to prepare 10 strains. 7 Each / mL spore suspension was inserted into the enzyme production medium (50mL / 250mL liquid volume) of the Erlenmeyer flask with 10% inoculum volum...

Embodiment 3

[0049] 1) Preliminary screening: The spore freeze-dried powder after space mutagenesis was gradiently diluted with sterile saline to make the spore concentration 10 3 cells / mL, take 0.05mL of the diluted sample, spread it on sterile cellulose-Congo red screening medium (121°C, sterilize for 30min), culture at 30°C for 48h, and select the transparent circle / colonies with larger diameter 1000 bacterial strains; wherein, the composition of the cellulose-Congo red screening medium is uniform: microcrystalline cellulose 10g, glucose 2g, agar 18g, Congo red 0.2g, Mandels inorganic nutrient salt 1000mL, pH value natural;

[0050] 2) Re-screening: The 1,000 strains selected in step 1) were inserted into the PDA slant medium, and the spores were covered with the slant, and the spores were washed with sterile saline to prepare 10 strains. 7 Each / mL spore suspension was inserted into the enzyme production medium (50mL / 250mL liquid volume) of the Erlenmeyer flask with 10% inoculum volume ...

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Abstract

The invention provides a trichoderma reesei strain capable of producing cellulase with high yield through space mutation, belonging to the technical field of microorganism mutation. According to the invention, trichoderma reesei Rw-X1 strain is preferably selected to be carried by Shenzhou IX to be subjected to space mutation and trichoderma reesei TG-C521 strain capable of producing cellulase with high yield can be carefully screened and cultured; the strain is collected in China General Microbiological Culture Collection Center on August 25, 2014 and has a collection Number of CGMCCNo.9537. According to the invention, the genetic character of the strain is changed by virtue of a special environment so that a strain with normal distributed mutation and stable genetic transformation can be screened out preferably; and the strain is used for producing cellulose and the filter paper activity of the strain is 25.83-34.49% higher than that of the trichoderma reesei Rw-X1 strain; the strain is friendly to the environment and is free of pollution, thus having a wide industrial application prospect.

Description

technical field [0001] The invention belongs to the technical field of microorganism mutation, and in particular relates to a Trichoderma reesei strain with high cellulase-producing space mutagenesis. Background technique [0002] Since the beginning of the 21st century, human beings are facing more and more severe challenges in energy, resources, and the environment. Cellulose is the largest renewable organic resource on the earth. Using these cheap and renewable plant cellulose raw materials to produce bio-based products and biomass energy is very beneficial to the sustainable development of human beings. Cellulose is the main component of plant cell walls, and is the most widely distributed and most abundant polysaccharide in nature, accounting for more than 50% of the carbon content in the plant kingdom. In my country, hundreds of millions of tons of crop stalks are burned every year, which not only pollutes the environment but also wastes resources. At present, the me...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N13/00C12N9/42C12R1/885
CPCC12N9/2437C12N13/00C12N1/145C12R2001/885
Inventor 王林风阎振丽赵子高吕世锋
Owner 河南天冠纤维乙醇有限公司
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