Secondary screening method of strain with high yield of lignocellulose degrading enzyme

A technology for lignocellulose and degrading enzymes, applied in fungi and other directions, can solve the problems of uneconomical practicality, high price, complicated operation steps, etc., and achieve the effects of strong industrialization pertinence, simple operation and high efficiency

Inactive Publication Date: 2015-04-22
熊鹏
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the current report, soluble sorbose (Sorbose), lactose (Lactose) and sophorose (Sophorose), and some insoluble carbon sources such as pure microcrystalline cellulose, xylan, pretreated lignocellulose, bovine Feces and corn fiber can induce microorganisms to produce enzymes, but in industrial production, these raw materials are expensive and not economical and practical. Therefore, only the inducers of industrial raw materials can be used to induce enzyme production, which can be more targeted in production
[0006] The general screening is to determine the cellulase activity or the ability of the microorganism to degrade cellulose by the size of the transparent circle formed by the microorganism on the plate culture medium with cellulose as the carbon source. This method needs to combine the pigment with the cellulose in advance, or The dyeing and decolorization step is carried out after microbial cultivation. If the staining is performed after microbial cultivation, the strains need to be backed up, which is very cumbersome.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1: Re-screen lignocellulose-degrading enzyme strains according to the following steps

[0024] (1) Basic culture medium: according to the mass of the metal ion solution, crushed filter paper containing 0.1% of its mass, 0.5% agar, and 0.1% ammonium sulfate (NH4) 2 SO 4 , 0.05% potassium dihydrogen phosphate KH 2 PO 4 , 0.01% corn steep liquor in a container, sterilized at 121°C for 30 minutes, then taken out and shaken well, and when the temperature was cooled to 60°C, metal ion solution was added to obtain the base layer culture medium; then the base layer culture medium was pipetted The liquid gun is dispensed into 20 mm of sterilized test tubes, placed in cold water to solidify quickly, the filter paper does not settle, and there are no air bubbles on the surface, and it is ready for use after solidification; wherein, the preparation method of the metal ionic liquid is as follows: first, prepare high-concentration metal ion Liquid, the concentration of...

Embodiment 2

[0028] Embodiment 2: Re-screen lignocellulose-degrading enzyme strains according to the following steps

[0029] (1) Basic medium: according to the mass of the metal ion solution, crushed filter paper containing 2.55% of its mass, 1.75% agar, 0.55% ammonium sulfate (NH4) 2 SO 4 , 0.275% potassium dihydrogen phosphate KH 2 PO 4 , 0.105% corn steep liquor was placed in a container, sterilized at 125°C for 25 minutes, then taken out and shaken well, and when the temperature was cooled to 75°C, metal ion solution was added to obtain the base layer culture medium; then the base layer culture medium was pipetted The liquid gun is dispensed into a sterilized test tube of 40 mm, placed in cold water to solidify quickly, the filter paper does not settle, and the surface has no air bubbles, and it is ready for use after solidification; wherein, the preparation method of the metal ionic liquid is as follows: first, prepare a high-concentration metal ion Liquid, the concentration of t...

Embodiment 3

[0033] Embodiment 3: Re-screen lignocellulose-degrading enzyme strains according to the following steps

[0034] (1) Basic culture medium: according to the mass of the metal ion solution, crushed filter paper containing 5% of its mass, 3% agar, 1% ammonium sulfate (NH4) 2 SO 4 , 0.5% potassium dihydrogen phosphate KH 2 PO 4 , 0.2% corn steep liquor in a container, sterilized at 128°C for 20 minutes, then taken out and shaken well, and when the temperature was cooled to 90°C, metal ion solution was added to obtain the base layer culture medium; then the base layer culture medium was pipetted The liquid gun is dispensed into a sterilized test tube of 60mm, placed in cold water to solidify quickly, the filter paper does not settle, and the surface has no air bubbles, and it is ready for use after solidification; wherein, the preparation method of the metal ionic liquid is as follows: first, prepare a high-concentration metal ion Liquid, the concentration of high-concentration...

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Abstract

The invention discloses a secondary screening method of strain with high yield of lignocellulose degrading enzyme. The method comprises (1) preparing basal culture medium; (2) preparing surface culture medium on the basal culture medium; (3) inoculating to-be-screened strain on the surface culture medium in test tube, and culturing in culture box; and (4) selecting the strains with vigorous growth, formation or no formation of spores, and relatively long transparent layer under black or other background observation as high-yield strain. Compared with traditional screening methods, the inventive secondary screening method has convenient and easy operation, stronger industrial pertinence, and effectiveness for screening high-yield strain adaptable to industrial culture medium.

Description

technical field [0001] The invention relates to a screening method for bacterial strains, in particular to a re-screening method for high-yielding lignocellulose-degrading enzyme strains. Background technique [0002] Lignocellulose is a natural and environmentally friendly resource with large reserves and renewable capacity in the world. The annual output of lignocellulose in the world is about 75 billion tons per year. In the modern society where environmental pollution and fossil energy are depleted day by day, the use of lignocellulose resources to replace fossil energy has become an urgent problem to be solved. The degradation of lignocellulose is the biggest difficulty in the utilization of lignocellulose resources. Countries around the world have invested huge human and financial resources in the research of lignocellulose degradation, through microbial screening, mutagenesis, enzyme purification, gene cloning and expression, metagenomics, metagenomics and other meth...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14
Inventor 熊鹏周玉珍吕睿瑞熊涛
Owner 熊鹏
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