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Esterase, encoding genes and use thereof

A technology of encoding genes and genes, which is applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve the problems of unseparated culture of microorganisms, difficulties in the isolation and cultivation of extremophiles, restrictions on the development and application of esterase genes of extremophiles, and achieve Effect of good activity and stability

Inactive Publication Date: 2009-01-28
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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Problems solved by technology

However, there are certain difficulties in the isolation and cultivation of extremophiles, which limit the development and application of extremophile esterase genes in special environments.
[0006] Studies have shown that currently the cultivable microorganisms in the environment only account for 1% of the total amount, and 99% of the microorganisms have not been isolated and cultured (Manuel Ferrer et al. Current Opinion in Biotechnology. 2005, 16(6): 588-593)

Method used

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  • Esterase, encoding genes and use thereof
  • Esterase, encoding genes and use thereof
  • Esterase, encoding genes and use thereof

Examples

Experimental program
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Effect test

Embodiment 2

[0039] Example 2, the acquisition of EstB gene and its functional verification

[0040] 1) Acquisition of EstB gene

[0041] Artificially synthesize the DNA molecule of sequence 1 in the sequence list, and add NdeI recognition sequence and HindIII recognition sequence at both ends of the sequence respectively. The plasmid pET28a was ligated overnight at 16°C under the action of T4 DNA ligase to obtain the recombinant plasmid pEEstB, image 3 It is a schematic diagram of the construction of the recombinant plasmid pEEstB. The recombinant plasmid pEEstB is transformed into competent Escherichia coli BL21, spread on LB medium containing 50 μl / ml kanamycin, and cultivated at 37°C for 16 hours. The positive clones were screened by colony PCR, and the obtained positive clones were named BL21EstB. Competent Escherichia coli BL21 was transformed with vector pET28a, and the recombinant strain BL21-28a obtained was used as a control.

[0042] Single colonies of recombinant bacteria B...

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Abstract

The invention discloses an esterase and coding gene and application thereof; the esterase disclosed by the invention is the protein of the following (a) or (b): (a) amino acid sequence thereof is classified as sequence 2 in a sequence list; (b) the amino acid sequence of the sequence 2 in the sequence list go through the substitution and / or deletion and / or addition of one or a plurality of amino acid residues and can catalyze and hydrolyze the protein of carboxylic ester, and the protein is derived from the (a). The invention also discloses a gene for encoding the esterase and the esterase of the invention is applicable to the industries of chemicals, food, biotransformation and medicine and other related industries.

Description

technical field [0001] The invention relates to an esterase, its coding gene and application. Background technique [0002] Esterase (EC 3.1.1.1) is a general term for all enzymes that can catalyze the hydrolysis of carboxylic esters, also known as carboxylesterase. It acts on both aliphatic and aromatic esters, and widely exists in animal tissues, plant seeds and microorganisms (KE Jaeger, et al. Trends in Biotechnology, 1998, 16(9): 396-403). [0003] As an important industrial enzyme, esterase is mainly used in catalytic reactions such as ester synthesis, lactone synthesis, transesterification, polypeptide synthesis, conversion and resolution of stereoisomers, etc. Esterase catalysis does not require coenzymes and has mild reaction conditions, The method has the advantages of simple method, high catalytic activity, strong selectivity, easy separation and recovery of products, and the like. Therefore be widely used in fields such as food processing, novel biomaterial, bi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/18C12N15/55C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21C12N15/11
Inventor 朱宝利胡永飞张立新
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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