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Method for screening cellulase producing strains and method for producing cellulase by means of fermentation

A screening method and cellulase technology are applied in the field of cellulase-producing strain screening and fermentation production of cellulase, and can solve the problems of high cellulase production cost, long cellulase fermentation time, and low fermentation enzyme activity. , to achieve the effect of not easy to contaminate bacteria, easy to pure culture, and high enzyme production rate

Inactive Publication Date: 2016-06-08
ANHUI UNIVERSITY OF TECHNOLOGY AND SCIENCE
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The problem of high production cost of cellulase is mainly manifested in the following two aspects: First, the fermentation enzyme activity of the strain producing cellulase is relatively low, which has always been an urgent problem to be solved in large-scale production, and it is also a long-term restriction on the mass production of cellulase. And the main factors that limit its application; second, the currently used filamentous fungi make cellulase fermentation time generally longer due to factors such as strain characteristics and culture conditions, and the cost is too high
At present, the filamentous fungi for cellulase production and research mainly include Trichoderma, Aspergillus and Penicillium. The fermentation cycle is generally more than 6 days, which has the characteristics of long time consumption and low efficiency.

Method used

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  • Method for screening cellulase producing strains and method for producing cellulase by means of fermentation
  • Method for screening cellulase producing strains and method for producing cellulase by means of fermentation
  • Method for screening cellulase producing strains and method for producing cellulase by means of fermentation

Examples

Experimental program
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Effect test

Embodiment 1

[0039] A screening method for producing cellulase strains, the screening method may further comprise the steps:

[0040] Primary screening:

[0041] Take 1g of the material containing the target strain (Huangshan ecological forest rotting wood) and pulverize; add 10mL of normal saline and shake for 1h. All the above suspensions were added to 200mL PDA medium, 30°C, 180rpm shaking culture for 12h;

[0042] The above culture was diluted 10-fold to 10-fold with normal saline -3 、10 -4 、10 -5, and spread them on the PDA plate medium respectively, and cultivate them statically at 30°C until colonies appear. Select a typical single colony and transfer it to the PDA slant, and make slices for purity inspection at the same time to obtain pure cultures; spot the pure cultures To the primary screening medium, the composition of the primary screening medium is: each L medium contains: CMC-Na5-10g; glucose 3-5g; agar 10-20g; MandelS inorganic nutrient salt 1000mL, cultured at 28°C for...

Embodiment 2

[0062] A screening method for producing cellulase strains, the screening method may further comprise the steps:

[0063] Others are the same as in Example 1, except that the composition of the primary screening medium is as follows: every L medium contains: CMC-Na 5g; Glucose 3g; Agar 10g; MandelS inorganic nutrient salt 1000mL;

[0064] The composition of the hyperosmotic primary screening medium plate is: each L medium contains: CMC-Na5g; glucose 3g; agar 10g; MandelS inorganic nutrient salt 1000mL; KCl0.6mol;

[0065] The composition of the liquid fermentation medium is: each L medium contains: 5g of 80-120 mesh corn stalks, 80g of 5% bran extract juice, (NH 4 ) 2 SO 4 20g, KH 2 PO 4 5g, MgSO 4 ·7H 2 O5g, CaCl 2 10g, Tween800.15g, trace elements: FeSO 4 ·7H 2 O0.003g, MnSO 4 ·H 2 O0.001g, ZnSO 4 ·7H 2 O0.001g, CoCl 2 ·6H 2 O0.001g, pH5.0.

Embodiment 3

[0067] A screening method for producing cellulase strains, the screening method may further comprise the steps:

[0068] Others are the same as in Example 1, except that the composition of the primary screening medium is: each L medium contains: CMC-Na 10g; Glucose 5g; Agar 20g; MandelS inorganic nutrient salt 1000mL;

[0069] The composition of the hyperosmotic primary screening medium plate is: each L medium contains: CMC-Na10g; glucose 5g; agar 20g; MandelS inorganic nutrient salt 1000mL; KCl0.6mol;

[0070] The composition of liquid fermentation medium is: every L medium contains: 80~120 order rice straw 10g, 5% bran extract juice 100g, (NH 4 ) 2 SO 4 30g, KH 2 PO 4 10g, MgSO 4 ·7H 2 O10g, CaCl 2 20g, Tween800.25g, trace elements: FeSO 4 ·7H 2 O0.005g, MnSO 4 ·H 2 O0.00156g, ZnSO 4 ·7H 2 O0.0014g, CoCl 2 ·6H 2 O0.002g, pH5.0.

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Abstract

The invention discloses a method for screening cellulase producing strains and a method for producing cellulase by means of fermentation. The cellulase producing strains which are named as TP-02 are acquired from rotten wood in ecological forest in the Huangshang Mountain by means of primary screening, protoplast preparation, ultraviolet-irradiation mutation breeding and secondary screening. The methods have the advantages that the cellulase producing strains are high in cellulase producing capacity, bean pulp, ammonium sulfate, urea or ammonium chloride can be used as nitrogen sources, bran juice, lactose, starch sugar, rice straw or corn straw can be used as carbon sources, the nitrogen sources and the carbon sources are subjected to liquid-state fermentation to obtain the cellulase producing strains, the enzyme activity of filter paper can reach 16.98 IU / mL after the cellulase producing strains are cultivated for 72-84 hours, the enzyme activity of incision enzymes can reach 41.33 IU / mL, the enzyme activity of excision enzymes can reach 3.65 IU / mL, and the enzyme activity of beta-glucanase can reach 15.98 IU / mL.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a screening method for cellulase-producing strains and a method for fermenting and producing cellulase. Background technique [0002] Lignocellulosic substances are the most widely distributed and abundantly stored substances on the earth, and they are also the cheapest renewable resources. Cellulase is a general term for a class of multi-component enzymes that can degrade cellulose into glucose. They work synergistically to degrade cellulose to produce oligosaccharides and cellodisaccharides, and finally hydrolyze them into monosaccharides such as glucose and xylose. In turn, alcohol, food, single-cell protein, pharmaceuticals and other chemical and chemical raw materials can be further produced. Therefore, cellulase has good application prospects in fuel ethanol production, papermaking, feed processing and other industries. [0003] Cellulase-producing bacteria includ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/01C12N13/00C12N9/42C12R1/845
CPCC12N15/01C12N9/2437C12N13/00
Inventor 汤斌张庆庆汤文晶李松
Owner ANHUI UNIVERSITY OF TECHNOLOGY AND SCIENCE
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