Trichoderma harzianum strain and use thereof

A technology for strains and lignin, applied in fungi, oxidoreductases, microorganisms, etc., can solve the problems of high production cost of ligninase, expanded culture, slow growth of bacteria, etc., and achieve broad industrial and agricultural applications and market prospects, fermentation Simple process, rapid growth and reproduction effect

Inactive Publication Date: 2009-08-12
CHINA AGRI UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In the traditional papermaking industry, lignin has always been separated and discarded as an obstacle to obtain papermaking fibers. Because the traditional method of separating lignin is carried out at high temperature with chemical reagents such as acid and alkali, resulting in The high cost and environmental pollution problems are the problems that have plagued the paper industry
[0007] In recent decades, some research progress has been made in ligninase and lignin biodegradation, but due to the high production cost of ligninase, it has been difficu...

Method used

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  • Trichoderma harzianum strain and use thereof
  • Trichoderma harzianum strain and use thereof
  • Trichoderma harzianum strain and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Isolation and Screening of Trichoderma harzianum Rifai WRF-2

[0042] PDA comprehensive medium: potato 200g, glucose 20g, peptone 10g, KH 2 PO 4 3g, MgSO 4 1.5g, V B1 10mg, 20g agar, 1000ml water, natural pH (do not control pH value). Peel fresh potatoes and cut into small cubes with a side length of 0.5cm, add 1000ml of water, boil and then heat for 10 minutes, then filter with four layers of gauze to obtain potato nutrient solution, add V B1 For other ingredients, add water to 1000ml, sterilize at 121°C for 15min, V B1 Separately make a solution and filter it with a 0.22 μm microporous membrane, then add it to make the final concentration 10 mg / L, mix it and spread it on a plate or make a slant medium.

[0043] Aniline blue (Azure-B) decolorization medium: yeast extract powder 10g, glucose 20g, aniline blue 0.1g, agar 18g, water 1000ml, pH natural, sterilized at 121°C for 15min. Plate after sterilization.

[0044] The isolation and screening steps of Trichod...

Embodiment 2

[0061] Fermentation of Trichoderma harzianum Rifai WRF-2

[0062] Utilize Trichoderma harzianum Rifai (Trichoderma harzianum Rifai) WRF-2 to produce lignin peroxidase (LiP) and / or manganese peroxidase, comprising the following steps:

[0063] 1. Spore culture and sporulation medium of Trichoderma harzianum Rifai WRF-2: 10 g of yeast extract powder, 20 g of glucose, 1000 ml of water, natural pH, sterilized at 121°C for 15 minutes.

[0064] Use an inoculation shovel to dig out a piece of mycelium with a side length of about 0.5 cm from the slope and inoculate it on a sporulation medium plate, and culture it at 30 ° C for 5 to 7 days to produce a large number of green spores.

[0065] 2. Production of lignin degrading enzymes by fermentation

[0066] Trace element solution: MgSO 4 ·7H 2O 3g, glycine 1.5g, MnSO 4 0.5g, NaCl 1g, FeSO 4 ·7H 2 O 0.1g, CoSO 4 0.1g, CaCl 2 0.08g, ZnSO 4 0.1g, CuSO 4 ·5H 2 O 0.01g, AlK(SO 4 ) 2 0.01g, H 3 BO 3 0.01g, NaMoO 4 0.01g, ...

Embodiment 3

[0074] Fermentation Production of Trichoderma harzianum Rifai WRF-2

[0075] Utilize Trichoderma harzianum Rifai (Trichoderma harzianum Rifai) WRF-2 to produce lignin peroxidase (LiP) and / or manganese peroxidase, comprising the following steps:

[0076] 1. The spore culture of Trichoderma harzianum Rifai (Trichoderma harzianum Rifai) WRF-2 is the same as that in Example 2.

[0077] 2. Production of lignin degrading enzymes by fermentation

[0078] The formulation of the enzyme-producing medium and the assay method of LiP and MnP enzyme activities are the same as in Example 2. Inoculate the spore suspension into 50 mL of enzyme-producing medium according to the method and concentration of Example 2, and cultivate for 3 days at 30° C. at 200 rpm. After the fermentation, the activity of LiP in the fermentation broth was determined to be 1908 U / L, and the activity of MnP was 3550 U / L. .

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Abstract

The invention relates to Trichoderma harzianum Rifai WRF-2 and use thereof. The bacterium strain was preserved in China General Microbiological Culture Collection Center on January 21st, 2009, with a preservation number of CGMCC No.2870. The bacterium belongs to deuteromycetes and is a lower fungus that is short in culture period and quick in growing and propagating speed and produces a large amount of uniform culture liquid for lignin degrading enzyme-generating bacteria in a short time compared with a major part of basidiomycetes to which whiterot fungi belong. The invention also provides a method for culturing and producing lignin peroxydase (Lip) and manganese peroxidase (MnP) by the fermentation of the bacterial strain in a liquid enzyme-generating culture medium, which can produce high-acitivity LiP and MnP in a short time, and is simple in fermentation process, stable, low in cost, and high in yield.

Description

technical field [0001] The invention relates to the field of microorganisms and applications thereof, in particular to a Trichoderma harzianum strain and applications thereof. Background technique [0002] Lignin is an important part of plant cell walls. Lignin accumulates in large quantities on the earth every year through plant photosynthesis. In nature, lignin is rich in content and is a natural polymer compound second only to cellulose. It is estimated that the global annual yields about 6 x 10 14 t lignin. At the same time, lignin is also degraded by microorganisms, participates in the carbon cycle on the earth, and plays an important role in it. [0003] Lignin is the product of plant tissue evolution, which endows plants with the ability to protect themselves and resist the decay of microorganisms. It is one of the most difficult biodegradable substances in natural organic matter. In nature, the degradation of lignin is a relatively slow process, which is the rate-...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12N9/02C12R1/885
Inventor 吴薇路明李桂英顿宝庆高振江
Owner CHINA AGRI UNIV
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