A gene-knockout Escherichia coli beneficial to extracellular secretion of recombinant protein and its application

A technology of Escherichia coli and gene knockout, which is applied in application, genetic engineering, plant gene improvement, etc., can solve the problems of low extracellular expression and increase downstream costs, and achieve enhanced extracellular secretion, low cost, and improved yield and purity Effect
CN104845926BInactive Publication Date: 2017-12-22NANJING TECH UNIV
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
NANJING TECH UNIV
Publication Date
2017-12-22
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention provides a gene-knockout Escherichia coli beneficial to the extracellular secretion of recombinant proteins and its application, and relates to the field of genetic genome transformation of Escherichia coli. The gene-knockout Escherichia coli which is beneficial to the extracellular secretion of the recombinant protein is obtained by knocking out the genes related to lipopolysaccharide synthesis in the Escherichia coli. The present invention also provides a construction method of the gene-knockout Escherichia coli and an application of the gene-knockout Escherichia coli in preparing extracellular soluble proteins. The preparation method of the gene-knockout Escherichia coli of the present invention is simple, high in efficiency and low in cost, and the permeability of the outer membrane of the cell is increased after the gene knockout of lipopolysaccharide synthesis is knocked out, thereby significantly enhancing the extracellular secretion of the recombinant protein.
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Description

technical field

[0001] The invention relates to the field of genetically modified Escherichia coli, in particular to a gene-knockout Escherichia coli beneficial to the extracellular secretion of recombinant proteins and its application. Background technique

[0002] The E. coli system is suitable for the production of proteins that do not require post-translational modification. Since Escherichia coli lacks the cofactors required in the protein folding process, the intermediates tend to accumulate in large quantities to form inclusion body precipitates. In order to obtain active proteins, the inclusion bodies must undergo tedious processes such as renaturation. The fructosidase is expressed in the Escherichia coli system. The intracellular expression is large and inclusion bodies are easy to form, and the extracellular expression is very small, which significantly increases the downstream cost.

[0003] Lipopolysaccharide (LPS) is a unique structural and functional molecule...

Claims

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