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Method for primarily screening high-yield cellulase funguses

A technology of cellulase and lignocellulose, applied in the direction of fungi, etc., can solve the problems of lack of pertinence and achieve the effect of strong industrialization pertinence

Inactive Publication Date: 2012-07-18
熊鹏
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the traditional laboratory screening formula containing pure cellulose lacks pertinence for the screening of relatively small differences in enzyme activity and the selective selection of excellent strains that use lignocellulose as the carbon source of the medium for fermentation. limitation

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1: Preliminary screening of cellulase fungus according to the following steps

[0022](1) Culture medium: First, according to the mass of the metal ionic liquid, crushed dry pretreated lignocellulose with a size of 50 mesh or more containing 0.1-5% of the mass, 0.5-3% agar, 0.1-1 % Ammonium Sulfate (NH 4 ) 2 SO4, 0.05-0.5% potassium dihydrogen phosphate KH 2 PO 4 , 0.05-0.5% (w / v) TritonX-100 is placed in a container, sterilized at 121-128°C for 20-30 minutes, then taken out and shaken well, when the temperature cools down to 60-90°C, add metal ionic liquid, Obtain the culture medium; then, pour the culture medium into a sterilized plate, and set aside after solidification; wherein, the preparation method of the metal ion liquid is as follows: first, prepare the metal ion concentrate, which contains magnesium sulfate, Ferrous sulfate, zinc sulfate, manganese sulfate, cobalt chloride and calcium chloride have a concentration of 10-200 times that of the met...

Embodiment 2

[0024] Embodiment 2: Preliminary screening of cellulase fungus according to the following steps

[0025] (1) Culture medium: First, according to the mass of the metal ionic liquid, crushed dry pretreated lignocellulose with a size of 50 mesh or more containing 0.1-5% of the mass, 0.5-3% agar, 0.1-1 % Ammonium Sulfate (NH 4 ) 2 SO4, 0.05-0.5% potassium dihydrogen phosphate KH 2 PO 4 , 0.05-0.5% (w / v) TritonX-100 is placed in a container, sterilized at 121-128°C for 20-30 minutes, then taken out and shaken well, when the temperature cools down to 60-90°C, add metal ionic liquid, Obtain the culture medium; then, pour the culture medium into a sterilized plate, and set aside after solidification; wherein, the preparation method of the metal ion liquid is as follows: first, prepare the metal ion concentrate, which contains magnesium sulfate, Ferrous sulfate, zinc sulfate, manganese sulfate, cobalt chloride and calcium chloride have a concentration of 10-200 times that of the me...

Embodiment 3

[0027] Embodiment 3: Preliminary screening of cellulase fungus according to the following steps

[0028] (1) Culture medium: First, according to the mass of the metal ionic liquid, crushed dry pretreated lignocellulose with a size of 50 mesh or more containing 0.1-5% of the mass, 0.5-3% agar, 0.1-1 % Ammonium Sulfate (NH 4 ) 2 SO4, 0.05-0.5% potassium dihydrogen phosphate KH 2 PO 4 , 0.05-0.5% (w / v) TritonX-100 is placed in a container, sterilized at 121-128°C for 20-30 minutes, then taken out and shaken well, when the temperature cools down to 60-90°C, add metal ionic liquid, Obtain the culture medium; then, pour the culture medium into a sterilized plate, and set aside after solidification; wherein, the preparation method of the metal ion liquid is as follows: first, prepare the metal ion concentrate, which contains magnesium sulfate, Ferrous sulfate, zinc sulfate, manganese sulfate, cobalt chloride and calcium chloride have a concentration of 10-200 times that of the me...

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PUM

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Abstract

The invention discloses a method for primarily screening high-yield cellulase funguses. The method comprises the following steps of: based on the mass of metal ionic liquid, placing 0.1-5 percent of pretreated lignocelluloses, 0.5-3 percent of agar, 0.1-1 percent of ammonium sulfate, 0.05-0.5 percent of monopotassium phosphate and 0.05-0.5 percent (w / v) of TritonX-100 in a container, sterilizing at 121-128 DEG C for 20-30 minutes, taking out and shaking up, and adding the metal ionic liquid when the temperature is reduced to 60-90 DEG C to obtain medium liquid; pouring the medium liquid into a sterilized plate, and condensing for later use; spreading strains to be screened on the plate, placing the plate in an incubator for 3-20 days, selecting according to the growth condition of each strain colony on the plate, and selecting strains with excellent enzyme producing activity. The method for primarily screening the high-yield cellulase strains has the advantages of low cost and high efficiency, and has stronger industrial pertinence as compared with the traditional screening method; in addition, by adopting the method, high-yield strains applicable to industrial medium can be effectively screened.

Description

technical field [0001] The invention relates to a screening method for bacterial strains, in particular to a primary screening method for high-yield cellulase fungi. Background technique [0002] Cellulase in a broad sense refers to the totality of cellulase secreted by microorganisms that can hydrolyze complex lignocellulose, and also includes auxiliary enzymes such as hemicellulase, Endoglucanases, EG, also known as endoglucanase , Endo-type cellulase, mainly acts on the non-crystalline region inside cellulose, randomly hydrolyzes β-1, 4 glycosidic bonds, shortens long-chain cellulose molecules, and produces a large number of small molecule cellulose with non-reducing ends ); Cellobiohydrolases, or CBH enzymes, also known as exoglucanase, exocellulase, cellobiohydrolase, (this type of enzyme acts on the reducing end of cellulose linear molecules to hydrolyze 1,4-β -D glycosidic bond, cut off a cellobiose molecule each time, so it is also called cellobiohydrolase); β-gluco...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14
Inventor 熊鹏周玉珍宇文伟刚熊涛
Owner 熊鹏