Space mutagenesis of Trichoderma reesei strain with high cellulase production

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

Inactive Publication Date: 2017-01-11
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 (v...

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 with high cellulase-producing space mutagenesis, and belongs to the technical field of microbial mutation. In the present invention, the Trichoderma reesei Rw‑X1 strain is preferably equipped with "Shenzhou No. 9" for space mutagenesis, and the Trichoderma reesei (Trichoderma reesei) TG‑C521 strain capable of high cellulase production is cultivated after careful screening. On August 25, 2014, it was deposited in the China General Microorganism Culture Collection Management Center, and the preservation number is CGMCC No.9537. The present invention uses a special environment to change the genetic shape of the bacterial species, and optimizes positive variation and stable genetic strains. This bacterial strain is used to produce cellulase, and its filter paper enzyme activity is 25.83% higher than that of Trichoderma reesei Rw‑X1 strain ~34.49%, environmental protection and pollution-free, has broad industrial application prospects.

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 Patents(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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