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Neutral endoglucanase as well as encoding gene and application thereof

An endoglucanase, neutral technology, applied in the fields of application, glycosylase, genetic engineering, etc., can solve problems such as limiting the application of endoglucanase, fading phenomenon, and narrow pH value edible range

Inactive Publication Date: 2015-06-03
SHENZHEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, the enzyme activity is low or has no activity, poor stability, narrow pH value edible range under neutral alkaline conditions, and the fading phenomenon will occur after textiles are treated under acidic conditions, which greatly limits the use of endoglucanases. Alkaline detergent, textile enzyme washing and finishing and other industrial applications

Method used

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  • Neutral endoglucanase as well as encoding gene and application thereof
  • Neutral endoglucanase as well as encoding gene and application thereof
  • Neutral endoglucanase as well as encoding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] (1) Materials

[0056] 1. Strains. Streptomyces sp. H31, a neutral endoglucanase-producing strain, was screened from Shenzhen mangrove soil. The strain preservation unit: China Center for Type Culture Collection (CCTCC), preservation time: January 5, 2015, preservation address: China. Wuhan. Wuhan University, preservation number: CCTCC NO: M 2015003. E.coli TOP10F' was purchased from Invitrogen;

[0057] 2. Carrier. The Escherichia coli cloning vector pMD-19T (simple) was purchased from Dalian Bao Biological Company, and the Escherichia coli expression vector pET-28a(+) (Novagen, KanR) was purchased from Novagen Company.

[0058] 3. Medium

[0059] (1) Selection medium: 10g / L CMC-Na (sodium carboxymethylcellulose), 5g / L yeast powder, 5g / L NaCl, 2g / L KH 2 PO 4 , 5g / L NH 4 NO 3 , 0.3g / L MgSO 4 , 1g / L (NH 4 ) 2 SO 4 , solid medium plus agar 15-20g / L, pH 9.0; culture temperature is 37°C.

[0060] (2) LB medium: tryptone 10g / L, yeast extract 5g / L, NaCl 10g / L, so...

Embodiment 2

[0114] (1) Materials

[0115] 1. Strains. Streptomyces sp. H31 strain (CCTCC No: M 2015003). The host bacteria E.coli BL21 Star(DE3) and E.coli TOP10F' were purchased from Invitrogen;

[0116] 2. Carrier. Escherichia coli expression vector pET-28a(+) (Novagen, Kan R ) was purchased from Novagen.

[0117] 3. Medium and buffer

[0118] (1) Selection medium: 10g / L CMC-Na (sodium carboxymethylcellulose), 5g / L yeast powder, 5g / L NaCl, 2g / L KH 2 PO 4 , 5g / L NH 4 NO 3 , 0.3g / L MgSO 4 , 1g / L (NH 4 ) 2 SO 4 , solid medium plus agar 15-20g / L, pH 9.0; culture temperature is 37°C.

[0119] (2) LB medium: tryptone 10g / L, yeast extract 5g / L, NaCl 10g / L, solid medium plus agar 15-20g / L, pH 9.0, autoclaved at 121°C for 20min.

[0120] (3) TE buffer: 10mmol / L Tris-Hcl, pH8.0, 1mmol / L EDTA, pH8.0.

[0121] (4) Alkaline lysis solution I, II, III (plasmid extraction): Alkaline lysis solution I: glucose 50mmol / L, Tris-HCl (pH 8.0) 25mmol / L, EDTA 10mmol / L. About 100mL per bottle, st...

Embodiment 3

[0180] Example 3 Research on Enzymatic Properties of Recombinant Neutral Endoglucanase H31

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Abstract

The invention discloses a neutral endoglucanase as well as an encoding gene and an application thereof. An H31 gene sequence is an intact open reading frame (ORF), wherein the open reading frame begins from an initiation codon ATG and is terminated by a termination codon TAG, and comprises 762 nucleotides; the nucleotide sequence is shown in SEQ ID No:1; an amino acid sequence of protein encoded by the neutral endoglucanase H31 gene is shown in SEQ ID No.2. The invention further discloses an application of the neutral endoglucanase H31 in industries such as washing and papermaking.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a neutral endoglucanase, its coding gene and application. Background technique [0002] Cellulose is the main component of plant fiber, accounting for about 30-50% of its dry weight. It is the most widely distributed natural carbohydrate and the most abundant and cheapest renewable resource on the earth. With the rapid increase of the world population, the rapid development of the global economy, the shortage of food and the reduction of non-renewable resources such as oil, coal and natural gas at an alarming rate, the development and utilization of renewable cellulose resources has attracted widespread attention around the world and has become A major topic of research in countries all over the world. The study of cellulase has opened up a broad way for this, especially in the past 10 years, with the rapid development of modern biotechnology, the application of gene rec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/42C12N15/56C12N15/70C12N1/21C11D3/386D21D1/02
CPCC11D3/38645C12N9/2437C12Y302/01004D21D1/02
Inventor 刘森林区晓阳邢苗陈伟钊
Owner SHENZHEN UNIV
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