Method for displaying human arginase1 on surfaces of escherichia coli

An arginase and Escherichia coli technology, applied in the surface field of human arginase-1, can solve the problems of carrying various viruses, long application time, complicated preparation process, etc. The effect of reducing synthesis cost and simplifying purification steps
CN105886491AInactive Publication Date: 2016-08-24HUBEI UNIV
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
HUBEI UNIV
Publication Date
2016-08-24
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention provides a method for displaying human arginase1 on surfaces of escherichia coli. The method comprises steps as follows: 1) recombinant plasmids for surface display of human arginase1 are constructed; 2) the recombinant plasmids are converted into escherichia coli competent cells, and genetic engineering strains are obtained; 3) the recombinant strains are subjected to shake-flask culture, and the display efficiency and enzyme activity of human arginase1 are detected; 4) L-Arginine (L-arginine, L-Arg) is efficiently converted by means of the recombinant strains for synthesis of L-Ornithine (L-ornithine, L-Orn). Human arginase1 is effectively displayed on surfaces of escherichia coli cells through improved Type V self-transport protein (Antigen 43), and industrial application of human arginase1 is realized more rapidly. By comparison with an original Type V self-transport protein (Antigen 43) display system, the display efficiency and the enzyme activity of human arginase1 are remarkably improved; by comparison with an existing chitin immobilization method, the synthesis cost is reduced, the technological process is shortened, and the purification procedures are simplified.
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Description

technical field

[0001] The present invention relates to the surface technology of human arginase-1 (Human Arginase 1), especially through the optimized autotransporter Antigen 43 surface display method, which is a new idea of ​​human arginase I display, New ways, new methods. Background technique

[0002] Human arginase-1 exists in human liver cells, participates in the urea metabolic cycle, and catalyzes the hydrolysis of L-Arg to generate L-Orn and urea. Arginase I deficiency can cause a series of related diseases, such as developmental delay, mental retardation, epilepsy and movement disorders. Human arginase-1 is also a potential clinical drug, which can be used to treat cancer.

[0003] Arginase is difficult to achieve high-efficiency soluble expression in Escherichia coli, and it is isolated and extracted from the liver, which is the main source of arginase, but there is a risk of carrying various viruses. Human arginase-1 has been secreted and expressed in Pichia p...

Claims

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