Cloning and amplified expression in Streptomyces lividans of a gene encoding extracellular β-lactamase from Streptomyces albus G

Abstract

A 4.9-kb DNA fragment containing the bla gene for the extracellular β-lactamase (BLA) of Streptomyces albus G was cloned in Streptomyces lividans using the conjugative, low-copy-number plasmid pIJ61 as vector. No expression of bla was observed when this DNA fragment was introduced into Escherichia coli HB101 on a plasmid vector. A 1.5-kb PstI- SstI fragment containing the bla gene was cloned in S. lividans on the non-conjugative, high-copy-number plasmid pIJ702. A tenfold higher yield of BLA was obtained from S. lividans carrying this plasmid than from S. albus G grown under optimal production conditions. The BLA from the clone reacts with β-iodopenicillanate according to a branched pathway which is characteristic of the original S. albus G BLA enzyme.