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Myrothecium verrucaria mutant strain T2901 and application thereof

A technology of verrucosia and mutant strains, applied in the field of microorganisms, can solve the problems of cellulose decline, carbohydrate loss, etc.

Inactive Publication Date: 2018-03-23
JILIN AGRICULTURAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, most lignin-degrading bacteria can only secrete one or two of the three lignin-degrading enzymes. For example, the model strain Phanerochaete chrysosporium that degrades lignin can only secrete lignin peroxidase and lignin peroxidase during fermentation. manganese peroxidase
[0007] At the same time, some white rot fungi degrade lignin and degrade cellulose and hemicellulose at the same time, resulting in the loss of carbohydrates. At the same time, the content of cellulose also dropped by 17.02%.

Method used

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  • Myrothecium verrucaria mutant strain T2901 and application thereof
  • Myrothecium verrucaria mutant strain T2901 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1 Isolation of bacterial strains

[0018] The soil in the forest area of ​​Changbai Mountain Nature Reserve was collected, and 1g of soil samples were taken in a 250mL Erlenmeyer flask filled with 99mL of sterile water, and shaken on a constant temperature oscillator at 30°C and 120r / min for 45min. After standing still, take the supernatant, and dilute according to the gradient multiple, and then use the dilution coating plate method to dilute the dilution of 10 -6 ~10 -8 The supernatant was transferred to a PDA plate, each dilution was repeated three times, and cultured in a 29°C incubator. After the flora grew out, they were repeatedly streaked and separated on the PDA plate, and after observing under a microscope to confirm that they were pure single colonies, they were inoculated on the preservation medium for preservation.

Embodiment 2

[0019] Example 2 Screening and Identification of Bacterial Strains

[0020] (1) Morphological identification and screening

[0021] Inoculate the isolated Verrucous verrucosa strain on PDA solid medium. The mycelium of the strain presents white flocculent at first, and grows divergently towards the periphery. , the conidia initially appeared dark green, and after 8 days of culture, the color of the conidia continued to deepen, and glue spots appeared; after 10 days of culture, the colony was in the shape of concentric rings, the conidia showed black glue spots, and light spots appeared on the back of the colony. Brown radial folds.

[0022] Use a puncher to get a 1cm-diameter bacterial block, inoculate it on an aniline blue selective medium, and culture it in an aerobic incubator at 30°C for 10 days, observe the fading of aniline blue, and select a strain with a fast fading rate and strong ability as candidate strains.

[0023] At the same time, in order to directional scre...

Embodiment 3

[0031] Example 3 Mutation and screening of bacterial strains

[0032] The preparation of mutant strains mainly adopts the atmospheric pressure room temperature plasma mutagenesis (ARTP) method, and the specific process is as follows:

[0033]Place 3-5 glass beads on the surface of the colony, add 1 mL of sterile water to prepare a spore suspension. Then dilute and adjust the spore concentration to 10 7 / mL, take 10 μL of the diluted bacterial suspension and drop it on the ARTP slide for mutagenesis.

[0034] Irradiate with the ARTP mutagenesis breeding system at a distance of 4 mm, and irradiate for 0 s, 15 s, 30 s, 45 s, 60 s, 75 s, 90 s, and 120 s, respectively. Put the treated slides in a centrifuge tube filled with 1 mL of sterile water, shake After uniformity, dilute 10 -2 、10 -3 , 10 -4 、10 -5 、10 -6 、10 -7 6 gradients, each gradient coating 3 parallel, determine the optimal mutagenesis time is 75s.

[0035] Spread the diluted bacterial suspension evenly on the...

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Abstract

The invention discloses a Myrothecium verrucaria mutant strain T2901 with a collection number of CCTCC NO: M2017413. The strain T2901 obtained by separation and mutagenesis can efficiently degrade lignin in corn straw. The strain T2901 can secrete laccase, lignin peroxidase and manganese peroxidase related to lignin degradation. Studies on impact of microbial pretreatment on structure and composition of the corn straw show that the strain T2901 has no loss of saccharides like cellulose and hemicelluloses, cellulose is retained greatly, and a new path for increasing straw cellulose enzymolysisutilization rate is provided.

Description

technical field [0001] The invention relates to the field of microbes, in particular to the application of Myrocarina verrucosus in bioenergy. Background technique [0002] The shortage of chemical energy and the increasingly serious environmental pollution make people constantly seek renewable green energy. As a carbon-neutral green energy, lignocellulose has become a good green candidate resource because of its abundant, easy-to-obtain and low-cost advantages. Sugar platform-based biorefinery technology is an effective way to utilize lignocellulose. This process mainly converts corn stalks, wheat straw and other lignocellulosic resources into polysaccharides, and then continues to convert polysaccharides into other bio-based products through biological fermentation. Among them, the annual output of corn stalks is high, but the utilization rate still needs to be improved. Enzymatic hydrolysis efficiently converts cellulose in corn stover to polysaccharides. However, lig...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P19/14C12P19/02C12R1/645
CPCC12P19/02C12P19/14C12P2201/00C12N1/145C12R2001/645
Inventor 孙旸陈光苏瑛杰于潇潇王刚陈欢唐珊珊苟泽昌李艳丽苏玉春吴卓夫张斯童
Owner JILIN AGRICULTURAL UNIV
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