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Alkaline endo glucanase gene, recombinase and applications thereof

An endoglucanase and enzyme gene technology, which is applied to the alkaline endoglucanase gene and its recombinase and application fields, and can solve the problems of low enzyme activity, lack, and little research on properties.

Inactive Publication Date: 2008-05-21
SHENZHEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

With regard to the research level of relevant alkaline endoglucanase in China at present, main problem is: (1) enzyme activity is still relatively low, can not reach the requirement of industrialized production level at home; (2) to the purification of enzyme, There are still few studies on the properties of enzymes, especially the studies on the properties of enzymes in detergents, and there is a lack of enzymes with practical application value; (3) The research on enzyme genes is not deep enough
These have hindered the industrialized production application of alkaline endoglucanase

Method used

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  • Alkaline endo glucanase gene, recombinase and applications thereof
  • Alkaline endo glucanase gene, recombinase and applications thereof
  • Alkaline endo glucanase gene, recombinase and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] (1) Materials

[0043] 1. Strains

[0044] Alkaliphilic Bacillus sp. III-3 (CGMCC No. 2138). Escherichia coli E.coli Top10 was purchased from Invitrogen.

[0045] 2. Instrument

[0046] MyCycler TM The thermal cycler PCR instrument, DNA and protein electrophoresis system are products of Bio-rad; the spectrophotometer and micro-spectrophotometer are products of Pharmacia Biotech; the Thermomixer comfort temperature-controlled oscillator, pipette gun, and desktop centrifuge are products of Eppendorf; The constant temperature shaker and constant temperature water bath are products of WAGEN; the homogenizer and low-temperature high-speed centrifuge are products of BECKMAN and Sigma; the Dolphin-DOC imaging system is a product of US WEALTEC.

[0047] (2) Experimental methods and results

[0048] 1. Genomic DNA Extraction

[0049] Use the EZ-10 spin column Genomic DNA Minipreps Kit (For Beteria) kit to extract Bacillus sp.III-3 genomic DNA, and see a relatively obvious...

Embodiment 2

[0066] 1. Materials and methods

[0067] (1) Materials

[0068] 1. Strains

[0069]Alkaliphilic Bacillus sp.III-3 strain (CGMCC No.2138); host bacteria E.coli BL21Star(DE3), E.coli Top10F' were purchased from Invitrogen.

[0070] 2. Carrier

[0071] Escherichia coli expression vector pET-28a(+) (Novagen) containing Kan + The resistance markers, vector maps and multiple cloning sites are shown in Figure 2 and Figure 3, respectively.

[0072] 3. Reagents and kits

[0073] Restriction endonucleases, PCR product fragment purification kits, gel recovery kits, DNA molecular weight markers, T4 ligase are products of Bao Biological Company; one-step rapid preparation of competent cell kit (SSCS) is from Shanghai Sangong Bioengineering Co., Ltd. Products; protein molecular weight markers are products of Fermentas (MBI); other reagents are domestic analytical grade. PCR primer synthesis and DNA sequence determination were completed by Takara Company.

[0074] 4. Medium and buffer...

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Abstract

The invention discloses an alkaline endoglucanase gene. The gene sequence is a complete open reading frame, which begins with ATG start codon and concludes with TAA stop codon. The whole sequence comprises 2502 nucleic acids. The invention also discloses the application of the recombinant alkaline endoglucanase in detergent industry etc. The recombinant alkaline endoglucanase has good resistance for SDS, which is preserved in the detergent with 0.2 percent final concentration for ten minutes without significant activity loss of the residue enzyme. The recombinant alkaline endoglucanase suits for the detergent and textile industries etc.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to an alkaline endoglucanase gene, its recombination enzyme and application. Background technique [0002] Endoglucanase (β-1, 4-endoglucanase, Endoglucanase / Endo-1, 4-β-Glucanase, EC3.2.1.4) is an important component of cellulase enzyme system, which can The cellulose molecule is cut from the inside at the β-1,4 glycosidic bond. Cellulase is a general term for a group of enzymes that degrade cellulose to generate glucose. It is often not a single enzyme, but a multi-component enzyme system that acts synergistically. In addition to endoglucanase, it also includes exoglucanase. Glycanase (β-1,4-exoglucanase, Cellobiohydrolase, CBH, EC 3.2.1.91) and β-glucosidase (BG, β-Glucosidase, EC 3.2.1.21). According to the optimal pH value of the catalytic reaction, endoglucanase can be divided into acidic endoglucanase (optimum pH 3-5), neutral endoglucanase (optimum pH 6-8) and alkali Sexual e...

Claims

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Application Information

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IPC IPC(8): C12N15/56C12N9/42C12N15/09C11D3/386
Inventor 邢苗刘森林陈伟钊
Owner SHENZHEN UNIV
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