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Method for preparing cellulase through trichoderma viride high-efficiency fermentation

A technology of Trichoderma viride and cellulase, which is applied in the field of microbial culture in biochemistry, can solve the problems of low enzyme production efficiency, poor solid fermentation stability, unsatisfactory substrate induction effect, etc., and achieves the effect of increasing economic income.

Inactive Publication Date: 2015-07-08
ZHEJIANG SCI-TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the practical problems such as low enzyme production efficiency, poor solid fermentation stability, and unsatisfactory substrate induction effect in the current process of microbial fermentation to prepare cellulase, at the same time, resource utilization of my country's abundant and low-cost moso bamboo resources is used to prepare high-addition Value biological enzyme preparation, the purpose of the present invention is to provide a kind of method for efficient fermentation Trichoderma viride to prepare cellulase

Method used

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  • Method for preparing cellulase through trichoderma viride high-efficiency fermentation
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  • Method for preparing cellulase through trichoderma viride high-efficiency fermentation

Examples

Experimental program
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Effect test

Embodiment 1

[0018] 1) Dissolve the unactivated Trichoderma viride with sterile water in a sterile environment, inoculate the strain on the Chapman medium by plate streaking, and cultivate it in a constant temperature incubator at 24°C for 72 hours until it grows. A single colony is produced to obtain activated Trichoderma viride;

[0019] 2) Pick a single colony of the activated Trichoderma viride obtained in step 1) in a sterile environment, inoculate it in a liquid activation medium, and cultivate it in a constant temperature shaker at 24°C and 180 r / min for 48 hours to obtain an expanded culture (liquid activation medium formula: glucose 10g / L, peptone 1g / L, Tween800.2g / L, ammonium sulfate 1g / L, potassium dihydrogen phosphate 1g / L, magnesium sulfate 0.1g / L, chloride Calcium 0.1g / L, urea 0.4g / L, Mandels nutrient salt accounting for 10% of the total volume of liquid activation medium, PBS with pH=3 as solvent, 120℃ high pressure steam sterilization for 20min)

[0020] 3) Inoculate the s...

Embodiment 2

[0022] 1) Dissolve the unactivated Trichoderma viride in sterile water in a sterile environment, inoculate the strain on the Chapman medium by streak method, and cultivate in a constant temperature incubator at 28°C for 108h until growth. A single colony is produced to obtain activated Trichoderma viride;

[0023] 2) Pick a single colony of the activated Trichoderma viride obtained in step 1) in a sterile environment, inoculate it into a liquid activation medium, and cultivate it for 60 hours in a constant temperature shaker at 28°C and 220 r / min to obtain an expanded culture (liquid activation medium formula: glucose 15g / L, peptone 1.5g / L, Tween 800.3g / L, ammonium sulfate 1.5g / L, potassium dihydrogen phosphate 2g / L, magnesium sulfate 0.2g / L, Calcium chloride 0.2g / L, urea 0.6g / L, Mandels nutrient salt accounting for 15% of the total volume of liquid activation medium, PBS with pH=5 as solvent, 120℃ high pressure steam sterilization for 20min)

[0024] 3) Inoculate the seed li...

Embodiment 3

[0026] 1) Dissolve the unactivated Trichoderma viride in sterile water in a sterile environment, inoculate the strain on the Chapman medium by streak method, and cultivate in a constant temperature incubator at 30°C for 144h until growth A single colony is produced to obtain activated Trichoderma viride;

[0027] 2) Pick a single colony of the activated Trichoderma viride obtained in step 1) in a sterile environment, inoculate it into a liquid activation medium, and cultivate it in a constant temperature shaker at 30°C and 240 r / min for 72 hours to obtain an expanded culture (liquid activation medium formula: glucose 20g / L, peptone 2g / L, Tween800.3g / L, ammonium sulfate 1.5g / L, potassium dihydrogen phosphate 2g / L, magnesium sulfate 0.3g / L, chlorine Calcium 0.2g / L, urea 0.8g / L, Mandels nutrient salt accounting for 15% of the total volume of liquid activation medium, PBS with pH=5 as solvent, 120℃ high pressure steam sterilization for 20min)

[0028] 3) Inoculate the seed liquid...

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Abstract

The invention discloses a method for preparing cellulase through trichoderma viride high-efficiency fermentation. The key points of the method are that: trichoderma viride is inoculated on a Czapek medium and is activated; a single colony is selected and is inoculated in a liquid activation culture medium and is subjected to amplification culture, such that a seed liquid is obtained. The seed liquid is inoculated into a liquid enzyme-production culture medium according to a volume ratio of 1:10-20; 5-20g / L of bamboo powder with a specification of 40-60 meshes is adopted as a fermentation substrate; liquid fermentation is carried out for 72-120h, such that cellulase is prepared. The method is stable and highly efficient. The method is especially suitable for producing cellulase through trichoderma viride high-efficiency fermentation. According to the invention, the entire set of liquid fermentation process is obtained through optimization aiming at trichoderma viride physiological characteristics, such that cellulase with high enzyme activity can be obtained. Also, the bamboo raw material is used as a resource and as a cellulase induction substrate, and the biological enzyme preparation with high added value is prepared. The invention provides an innovative idea for utilizing bamboo resource with high value. With the method, Bamboo area farmers' economic income can be effectively improved. The invention has important significance.

Description

technical field [0001] The invention relates to a method for preparing cellulase, in particular to a method for efficiently fermenting Trichoderma viride to prepare cellulase, and belongs to the technical field of microbial culture in biochemistry. Background technique [0002] Cellulase is a general term for a group of multi-component enzymes that can hydrolyze the glucosidic bonds of cellulose molecular chains to convert them into glucose. Cut cellulase, β-glucosidase, etc. Cellulose requires the co-occurrence and synergistic action and co-catalysis of these enzymes to be completely degraded. Using cellulase to convert cellulose into food, energy and chemical materials has great practical significance for solving many resource and environmental problems faced by human beings. [0003] The current bottleneck hindering the widespread and efficient utilization of cellulase by human beings is the lack of high-producing bacteria of cellulase, and the process of fermentation e...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/42C12R1/885
CPCC12N9/2437
Inventor 姚菊明庄源张勇崔建梅
Owner ZHEJIANG SCI-TECH UNIV
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