Biological preparation method of chitin deacetylase

A technology for chitin deacetylase and biological preparation, which is applied in the field of acetylase preparation, can solve the problems of long time consumption and low yield of chitin deacetylase, and achieves the advantages of short production cycle, high yield and output, and low extraction cost. Effect

Inactive Publication Date: 2010-03-03
SHANGHAI OCEAN UNIV
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Problems solved by technology

[0014] In order to overcome the shortcomings of traditional biochemical technology, the time-consuming and low yield of chitin deacetylase separation and acquisition, which can only meet the needs of laboratories, the purpose of the present invention is to provide a chitin deacetylase that can be prepared quickly and in large quantities. Methods

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  • Biological preparation method of chitin deacetylase
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Embodiment Construction

[0040] In order to make the technical means, creative features, goals and effects of the present invention easy to understand, the present invention will be further described below in conjunction with specific illustrations. It should be understood that these embodiments are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods without specific conditions indicated in the following examples generally follow conventional methods, and techniques not specified in detail belong to conventional techniques.

[0041] The gene and cDNA cloning and gene recombination methods used in the present invention are not particularly limited, and conventional molecular cloning techniques in the field of molecular biology can be used. The main methods of the present invention include: starting strain screening, cDNA cloning, DNA recombination, prokaryotic expression and protein purification schemes.

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Abstract

The invention discloses a biological preparation method of chitin deacetylase derived from racemomucor, which comprises the following steps: (a) preparing a recombinant expression vector of the chitindeacetylase containing a gene sequence such as SEQ ID NO.1; (b) preparing a transformant containing the recombinant expression vector in the step (a); and (c) expressing and purifying the chitin deacetylase. In the invention, a gene specificity degenerate primer is designed, a reverse transcription and polymerase chain reaction and a rapid amplification technology of cDNA segment ends are adopted, and escherichia coli strains which can generate the chitin deacetylase are obtained by combining the DNA recombination method and utilizing the known pronucleus express vector. The invention provides a reliable source for biologically preparing the chitin deacetylase and has the advantages of short production period, low extraction cost, higher product enzyme yield, higher output, and the like.

Description

technical field [0001] The invention relates to the field of enzyme engineering of biotechnology, and more specifically relates to a method for preparing acetylase by obtaining engineering bacteria producing chitin deacetylase by means of genetic engineering. Background technique [0002] Chitin is a high molecular substance, its content in nature is second only to cellulose, and the global annual output of shrimp and crab shells is nearly 20 million tons. Chitin is insoluble in water, dilute acid, dilute alkali and other organic solvents, which greatly limits its application range. Currently the most widely used deacetylated derivative of chitin - chitosan. [0003] However, the process of deacetylating chitin into chitosan must have the conditions of concentrated alkali (at least 40% or more) and higher temperature (more than 90°C), which not only puts high demands on the production equipment, but also Encountered the difficult problem of "how to deal with the concentrat...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/70C12N9/80C12R1/19
Inventor 蒋霞云邹曙明
Owner SHANGHAI OCEAN UNIV
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