Construction and fermentation method of artificial strain with high yield of fengycin

Abstract

The invention provides a construction and fermentation method of an artificial strain with high yield of fengycin. The 4'-phosphoric acid pantetheinyl transferase gene sfp from bacillus amyloliquefaciens FZB42, a multi-effect factor gene degQ from bacillus subtilis 168 and a strong promoter P43 are integrated in series onto a genome of the bacillus subtilis 168 by utilizing CRISPR gene editing to construct a strain B.subtilis ds; an acetyl coenzyme A synthetase gene acs of Escherichia coli BL21, an acetyl coenzyme A carboxylase gene accACD of Salmonella enterica and a biotin ligase gene bisA of Corynebacterium glutamicum are connected in series to an expression vector pHT43, and the expression vector pHT43 is introduced into a fengycin synthesis strain B.subtilis ds, so that a fengycin high-yield strain B.subtilis dspabacd is constructed; the constructed engineering strain is fermented and cultured in a shake flask to produce the fengycin.

Description

technical field

[0001] The invention belongs to the technical field of microorganisms, and relates to the serial integration of the 4'-phosphopantetheinyl transferase gene sfp regulatory factor gene degQ into the genome of Bacillus subtilis 168, and the overexpression of the acyl-CoA synthetase gene acs, Biotin ligase gene birA and acetyl-CoA carboxylase gene accACD, a method for constructing a high-yield Fengyuansu strain. Background technique

[0002] Fengyuan element (fengycin) is synthesized by Bacillus through non-ribosomal peptide synthesis pathway. It is composed of a β-hydroxy fatty acid chain containing 16 to 18 carbon atoms and ten amino acid residues. The factor can have a good inhibitory effect on filamentous fungi. So far, the mechanism of the inhibitory effect of the factor on the fungus is still inconclusive. At present, the theory accepted by the academic circle is that the factor interacts with the phospholipid bilayer of the cell. It penetrates the phospho...

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