Egg-white lysozyme and preparation method thereof

A technology for egg white lysozyme and lysozyme, which is applied in the field of egg white lysozyme and its preparation, and achieves the effects of increasing copy number, efficient synthesis and high biological activity

Inactive Publication Date: 2015-01-07
TIANJIN SHENGJI GRP CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Since lysozyme is applied to this industry for the first time, there are very few reports on the application of lysozyme in livestock and poultry in the current market. Therefor

Method used

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  • Egg-white lysozyme and preparation method thereof
  • Egg-white lysozyme and preparation method thereof
  • Egg-white lysozyme and preparation method thereof

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Embodiment 1

[0033] A method for preparing egg white lysozyme, using gene synthesis technology to obtain the cDNA sequence of egg white lysozyme, using the E.coli expression system to efficiently express the egg white lysozyme lyz gene, and constructing recombinant Escherichia coli containing the target gene lyz , under the induction of IPTG, ferment and synthesize, and finally obtain egg white lysozyme through extraction and purification. The target gene used in the present invention is synthesized according to the egg white lysozyme protein sequence (ACCESSIONNUMBER: GI345100466) published by NCBI, and the host cell of the recombinant vector in the present invention is Escherichia coli BL21 (DE3). The constructed recombinant expression vector not only includes the DNA sequence encoding the enzyme, but also has the control elements required for expressing the gene.

[0034] The steps of the specific construction method are as follows:

[0035] 1. Synthesis of egg white lysozyme lyz gene ...

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Abstract

The invention provides egg-white lysozyme and a preparation method thereof. The coding sequence of polypeptide amino acid with protein activity of the egg-white lysozyme is a sequence show in a sequence table 400<1>. The preparation method comprises the following steps: (1) obtaining genes of the egg-white lysozyme; (2) constructing recombinant plasmids; (3) constructing genetic engineering strains; and (4) carrying out producing method of genetic engineering recombinant protein. The egg-white lysozyme has the characteristics of stability, purity and high bioactivity and can be widely applied multiple field, such as pharmacy and feed. According to the preparation method, the genetic engineering strains act on fermentatively synthesized lysozyme, by adopting constructed engineering strains, the copy number of enzyme genes can be greatly increased, the expression quantity of the enzyme is improved, and thus the efficient synthesis of the egg-white lysozyme is achieved and the preparation method of the egg-white lysozyme is not limited by the source of raw materials.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to egg white lysozyme and a preparation method thereof. Background technique [0002] Lysozyme (lysozyme) was first published by Nicolle (Nicolle) l907 report on the lytic factor of Bacillus subtilis. Two years later, Laschtscbenko pointed out that egg white has a strong antibacterial effect, which is the result of enzyme action. In 1922, Fleming discovered that human mucus, saliva, and tears also have strong bacteriolytic activity, and it was named lysozyme because of its bacteriolytic effect. Lysozyme is widely found in animals, plants and microorganisms. It is a natural antibacterial agent that can specifically act on cell wall N-acetylglucosamine (N-acetylglucosamine, NAG) and N-acetylmuramic acid (N-acetylmuramic acid). acid, NAM) between the β-1, 4 glycosidic bonds, destroy the peptidoglycan scaffold, make the microbial cells swell and crack under the action of intern...

Claims

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Application Information

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IPC IPC(8): C12N9/36C12N15/70
CPCC12N9/2462C12Y302/01017
Inventor 王连民刘珂飞荆玮
Owner TIANJIN SHENGJI GRP CO LTD
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