A method for constructing high-efficiency secretion and expression of transpeptidase sortase A in Escherichia coli

Abstract

The invention discloses a method for constructing Escherichia coli capable of efficiently secreting and expressing transpeptidase Sortase A, and belongs to the field of gene engineering. According tothe present invention, the efficient secretion and expression of SortaseA in Escherichia coli is successfully achieved, and the problems that the transpeptidase Sortase A cannot be secreted and expressed and the expression level of the transpeptidase Sortase A in cells is low in the existing report are solved; at the shake flask level, the yield of the obtained sortedaseA can achieve 90.2 mg / L, and the extracellular enzyme activity can achieve 34.2 U / mL; and the foundation is established for the subsequent industrial production of SortaseA.

Description

technical field

[0001] The invention relates to a method for constructing Escherichia coli to efficiently secrete and express transpeptidase Sortase A, belonging to the field of genetic engineering. Background technique

[0002] Sortase A is a class of transpeptidases that mediate the covalent binding of Gram-positive bacterial cell wall-anchored proteins to the cell wall. Its application in the field of biotechnology has attracted more and more attention in recent years. Since the first report of using Sortase A as a linking tool for proteins or polypeptides in 2004, the linking reaction involving Sortase A has rapidly become a research hotspot and has been widely used in biochemistry, proteomics, biomedicine, and bioengineering technologies, etc. field. But so far, there are few studies on the high-efficiency expression and industrial production of Sortase A, and the expression method of the enzyme is still mainly limited to the intracellular expression of Escherichia col...

Images