Cloning and Expression of the Gene Encoding Mannose Enzyme II of the Corynebacterium glutamicum Phosphoenolpyruvate-Dependent Phosphotransferase System in Escherichia coli

Abstract

The gene for mannose enzyme II of phosphoenolpyruvate-dependent phosphotransferase system from Corynebacterium glutamicum KCTC 1445 was cloned into Escherichia coli ZSC113 using plasmid pBR 322. The recombinant plasmid, designated pCTS3, contained 2.2 kb DNA fragment, and the physical map of the cloned DNA fragment was determined. The E. coli ptsM ptsG mutant transformed with pCTS3 restored glucose and mannose fermentation ability, and grew well on these sugars as the sole carbon source in the minimal medium. The transform ant harboring pCTS3 showed a PTS-mediated repression of growth on maltose by mannose analogue, 2-deoxyglucose. The specificity of the response to 2DG therefore indicates that the cloned DNA fragment carries mannose enzyme II gene.

Keywords

• phosphoenolpyruvate:glycose phosphotransferase system(PTS);
• mannose transport;
• mannose enzyme II;
• Corynebacterium glutamicum