Cloning and location of a gene governing lysine epsilon-aminotransferase, an enzyme initiating beta-lactam biosynthesis in Streptomyces spp

Author:

Madduri K1,Stuttard C1,Vining L C1

Affiliation:

1. Department of Microbiology, Dalhousie University, Halifax, Nova Scotia, Canada.

Abstract

In actinomycetes that produce beta-lactam antibiotics of the cephem type, lysine epsilon-aminotransferase is the initial enzyme in the conversion of lysine to alpha-aminoadipic acid. We used a two-stage process ("chromosome walking") to screen a lambda library of Streptomyces clavuligerus genomic DNA for fragments that expressed lysine epsilon-aminotransferase activity in S. lividans. Restriction analysis of the cloned DNA confirmed the location of the putative lat gene within the cluster of beta-lactam biosynthesis genes, roughly midway between pcbC, the structural gene for isopenicillin N synthetase, and the putative cefE gene encoding deacetoxycephalosporin C synthetase.

Publisher

American Society for Microbiology

Subject

Molecular Biology,Microbiology

Reference18 articles.

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2. Demain A. L. 1983. Biosynthesis of P-lactam antibiotics p. 189-228. In A. L. Demain and N. A. Solomon (ed.) Antibiotics containing the beta-lactam structure vol. 1 Springer Verlag New York.

3. Hopwood D. A. M. J. Bibb K. F. Chater T. Kieser C. J. Bruton H. M. Kieser D. J. Lydiate C. P. Smith J. M. Ward and H. Schrempf. 1985. Genetic manipulation of Streptomyces: a laboratory manual. John Innes Foundation Norwich England.

4. Plasmids, recombination and chromosome mapping in Streptomyces lividans 66;Hopwood D. A.;J. Gen. Microbiol.,1983

5. Jones G. H. 1989. Cloning of Streptomyces genes involved in antibiotic synthesis and its regulation. p. 49-73. In S. Shapiro (ed.) Regulation of secondary metabolism in actinomycetes. CRC Press Boca Raton Fla.

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