Trichoderma viride strain, and applications thereof

A technology of Trichoderma viride and strains, applied in the field of microorganisms, can solve the problems of cumbersome operation, limited bacterial tolerance, and low enzyme protein production, and achieve the effects of high enzymolytic activity, strong tolerance, and high enzymatic activity

Active Publication Date: 2016-06-08
CHINA PETROLEUM & CHEM CORP +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] Most of the currently obtained cellulase-producing strains directly use lignocellulosic raw materials as carbon sources and enzyme protein inducers, and the enzyme protein production is small, and the secreted cellulase is adaptable to enzymatic hydrolysis of pretreated lignocellulose substrates Not strong, used for enzymatic hydrolysis of lignocellulosic raw materials for pretreatment, the yield of glucose is not high
In order to improve the enzymatic hydrolysis effect of cellulase on pretreated lignocellulose, pretreated lignocellulose raw materials can be used as carbon source and inducer. , will release some inhibitors of microbial growth, including weak acid inhibitors (formic acid, acetic acid, levulinic acid, etc.), furfural inhibitors (furfural, 5-hydroxymethylfurfural) and phenolic inhibitors (vanillin, on Hydroquinone, 4-hydroxybenzoic acid), etc. These inhibitors, especially phenols, have a certain inhibitory effect on the production of cellulase and cellulose enzymolysis by the strain, so detoxification treatments such as neutralization and water washing are required. cumbersome
If the pretreated lignocellulosic raw material is directly used, the yield and activity of cellulase are not high due to the limited tolerance of the bacteria

Method used

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  • Trichoderma viride strain, and applications thereof
  • Trichoderma viride strain, and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024]The separation and screening of embodiment 1 Trichoderma viride F4

[0025] Trichoderma viride F4 of the present invention ( Trichodermaviride F4) was obtained by screening from the soil of the roots of pine trees on the top of the mountain in Fushun City, Liaoning Province. It was obtained by enrichment, wood block coating screening, filter paper coating screening, and Congo red-CMC double-layer plate screening. The specific screening process is:

[0026] (1) Dilute the obtained soil sample, suspend it in sterile water, add it to the fermentation medium, and incubate at 30°C for 72-84h;

[0027] (2) Take an appropriate amount of culture medium in step (1), press 10 －4 , 10 －5 , 10 －6 Dilute, spread on wood block, and incubate at 30°C for 72h;

[0028] (3) Select the one with the largest colony and inoculate it in the fermentation medium, and incubate at 30°C for 72-84 hours;

[0029] (4) Take an appropriate amount of culture medium, press 10 －4 , 10 －5 , 10 －6...

Embodiment 2

[0036] Embodiment 2 Trichoderma viride F4 liquid state fermentation produces cellulase

[0037] The above pretreated corn stalks containing inhibitors were used as carbon source and inducer to produce cellulase by liquid fermentation of Trichoderma viride F4. The specific process is as follows:

[0038] (1) Preparation of liquid fermentation medium: wheat bran 50g / L, pretreated corn straw 50g / L, microcrystalline cellulose 25g / L, yeast extract 20g / L, KH 2 PO 4 0.25g / L, CaCl 2 0.4g / L, MgSO 4 0.4g / L, ZnSO 4 ·7H 2 O6mg / L, MnSO 4 4H 2 O2.5mg / L, CoCl 2 2mg / L;

[0039] (2) Place the above culture medium in a 500mL Erlenmeyer flask, sterilize at 121°C for 30 minutes, inoculate Trichoderma viride F4 after cooling, incubate at a constant temperature of 30°C for 24 hours, and rotate at 200rpm to obtain seed liquid;

[0040] (3) Inoculate the above seed solution into the fermentation medium at an inoculation amount of 10% (v / v), and ferment at 30°C for 72 hours;

[0041] (4) Us...

Embodiment 3

[0046] Embodiment 3 Trichoderma viride F4 liquid state fermentation produces cellulase

[0047] The treatment process and operating conditions are the same as in Example 2, except that the prepared Trichoderma viride F4 seed liquid is inoculated into 5 identical mediums respectively, and after 24 hours of fermentation, 1.5g pagoda tree powder and sorbose are added respectively , cellobiose, isomaltose, gluconic acid, and cultured for another 72 hours. Three batches were repeated to detect the FPA, CMC enzyme activity, and β-glucosidase activity of different samples, as described in Table 1.

[0048] Table 1 Enzyme activity detection results of different samples (unit: IU / ml)

[0049]

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Abstract

The invention discloses a Trichoderma viride F4 strain, and applications thereof. The Trichoderma viride F4 strain is stored at China General Microbiological Culture Collection Center on 10th November 2008, and preservation number is CGMCC No.2736. When the bacterial strain is used for producing cellulase taking pretreated lignocellulose as a carbon source and an inducer, the bacterial strain is capable of tolerating a plurality of inhibitors brought by pretreated lignocellulose, and producing enzyme with high efficiency; the secreted cellulase is comprehensive in composition, enzyme activity is high, and enzymolysis adaptability on pretreated lignocellulose substrate is high.

Description

technical field [0001] The invention belongs to the technical field of microbes, and relates to a Trichoderma viride strain and its application, in particular to a Trichoderma viride strain with strong resistance to inhibitors and its application in the preparation of cellulase. Background technique [0002] Cellulose is the largest renewable organic resource in the world. The global biosynthesis of renewable cellulose reaches more than 100 billion tons every year, but it is rarely utilized. 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 industrializatio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N9/42C12P19/14C12P19/02C12R1/885
Inventor 唐开宇张全
Owner CHINA PETROLEUM & CHEM CORP
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