Trichoderma harzianum strain from Tibet for producing cellulase and application of trichoderma harzianum strain

A technology of cellulase and lignocellulose, applied in the field of microorganisms

Pending Publication Date: 2019-12-03
JALA GROUP CORPORATION +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the prior art, there are few reports about Trichoderma harzianum strains with high cellulase production and its application in lignocellulosic biomass degradation

Method used

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  • Trichoderma harzianum strain from Tibet for producing cellulase and application of trichoderma harzianum strain
  • Trichoderma harzianum strain from Tibet for producing cellulase and application of trichoderma harzianum strain
  • Trichoderma harzianum strain from Tibet for producing cellulase and application of trichoderma harzianum strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Screening and biological identification of Trichoderma harzianum LZ117

[0043] (1) Screening of Trichoderma harzianum LZ117

[0044] Put the plant and soil samples collected in different geographic locations in the Tibet plateau into 50mL sterile centrifuge tubes, add 20mL of sterilized pure water, vortex fully for 60s, and carry out gradient dilution of the sample suspension (10 2 After that, 100 μL of the diluted sample solution was taken and spread on Mandels solid medium with cellulose as the single carbon source, in which 1% (weight / volume) streptomycin was added to inhibit bacterial growth. After culturing at 25°C for 48 hours, observe the production of the transparent circle. As a result, it was found that among the 35 samples, more than 10 samples had cellulose degradation transparent circles in the coated flat plates. Scraping and diluting the spores of filamentous fungal strains capable of degrading cellulose selected from each plate (or picking t...

Embodiment 2

[0052] The growth state of embodiment 2 Trichoderma harzianum LZ117 in different carbon source media, different temperatures

[0053] Inoculate a small amount of spores or hyphae of Trichoderma harzianum LZ117 into MEA medium to produce spores, culture in an incubator at 28°C for 7-10 days, wash fresh conidia with sterile distilled water and filter them with sterilized 6-layer gauze to prepare spore suspension. Measure the concentration of the spore suspension through a hemocytometer and dilute to 10 7 individual / mL, then take 2 μL of spore suspension and inoculate them into the center of Mandels solid medium and MEA solid medium plates containing 2% (weight / volume) different carbon sources (such as glucose, lactose, cellulose), respectively, and place Cultivate at 4°C, 15°C, 20°C and 28°C for 5 days, and observe the growth state of bacteria in each medium. The results showed that the growth of LZ117 strain was significantly inhibited when cultured at 4°C under different car...

Embodiment 3

[0054] Example 3 Trichoderma harzianum LZ117 extracellular cellulase enzymatic properties

[0055] The filter paper enzyme activity of the extracellular cellulase of the LZ117 strain was measured by phosphate-citrate buffer solution of different pH (pH 3.0-7.0) at 50°C. Experiments show that the filter paper has the highest enzyme activity under the condition of pH 5.0, the results are shown in the attached figure 1 . At the same time, under the condition of pH 5.0, different enzyme reaction temperatures (30°C, 35°C, 40°C, 45°C, 50°C, 55°C and 60°C) were changed to measure the activity of extracellular cellulase filter paper. Experiments show that it has the highest filter paper enzyme activity at 50°C, the results are shown in figure 2 . Finally, the optimal temperature and pH value of the extracellular cellulase were determined to be 50°C and pH 5.0, respectively.

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Abstract

The invention discloses a trichoderma harzianum strain LZ117 from Tibet for producing cellulase. The preservation number of the trichoderma harzianum strain LZ117 is CGMCC No.17184, and the trichoderma harzianum strain LZ117 and a fermentation liquor thereof can be used for degrading lignocellulose biomass. A preparation method of the cellulase-containing fermentation liquor produced by the trichoderma harzianum strain LZ117 is also disclosed. According to the trichoderma harzianum strain LZ117 from Tibet for producing the cellulase, a new microbial resource is provided for degradation of thelignocellulose biomass; after the trichoderma harzianum strain LZ117 is cultured for 48 hours in a Mandels plate which adopts cellulose as a sole carbon source, obvious degradation transparent circlescan be shown; after culture is conducted in a 250 mL shake flask for about 120 hours, the fermentation enzyme liquor can achieve maximum filter paper activity, and high-activity cellulose degrading capability of the fermentation enzyme liquor is reflected; and the trichoderma harzianum strain LZ117 has high capability of degrading filter paper, preprocessed corn straw and preprocessed jerusalem artichoke straw, and accordingly has great application potentiality in the aspects of lignocellulose biomass resource development and utilization.

Description

technical field [0001] The invention relates to the technical field of microbes, in particular to a Trichoderma harzianum strain producing cellulase from Tibet and its application. Background technique [0002] Lignocellulosic biomass includes crop straw, forestry waste, urban greening waste, etc., and has the characteristics of low price, abundant resources and renewable. Cellulase mainly has three types of enzyme components, exocellulase, endocellulase and β-glucosidase, and these enzymes complete the degradation of lignocellulose through synergistic action. The cellulose and hemicellulose components of such biomass are degraded by lignocellulosic degrading enzymes, releasing fermentable reducing sugars that can be fermented by microorganisms to produce biofuels and biobased chemicals. The biorefining technology of lignocellulosic biomass can replace the existing technology of non-renewable petroleum, so it has attracted extensive attention of scholars at home and abroad....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N9/42C12R1/885
CPCC12N9/2437C12R2001/885C12N1/145
Inventor 李俊赵心清章漳李嘉祥张飞白凤武
Owner JALA GROUP CORPORATION
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