Characterization and complementation of a cephalosporin-deficient mutant of Streptomyces clavuligerus NRRL 3585

• Published in: Applied microbiology and biotechnology Vol. 32; no. 5; pp. 560 - 567
• Main Authors: PIRET, J, RESENDIZ, B, MAHRO, B, JIN-YOU ZHANG, SERPE, E, ROMERO, J, CONNORS, N, DEMAIN, A. L
• Format: Journal Article
• Language: English
• Published: Berlin Springer 01-02-1990
• Subjects: Bacterial Proteins - metabolism; Bacteriology; Biological and medical sciences; Biology of microorganisms of confirmed or potential industrial interest; Biotechnology; cephalosporins; Cephalosporins - biosynthesis; cloning; Cloning, Molecular; complementation; DNA; DNA, Bacterial - genetics; DNA, Bacterial - isolation & purification; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation, Bacterial - genetics; Genetic Complementation Test; hybridization analysis; Metabolism. Enzymes; Microbiology; Mission oriented research; mutants; Mutation; nucleotide sequence; Oxidoreductases - genetics; Peptide Synthases - genetics; Phenotype; Physiology and metabolism; Plasmids - genetics; Restriction Mapping; Streptomyces - enzymology; Streptomyces - genetics; Streptomyces - metabolism; Transformation, Bacterial; Molecular hybridization; Streptomycetaceae; Copy number; Metabolism; Actinomycetales; Characterization; Plasmid; Antibiotic; Cephalosporin derivatives; Enzymatic activity; Bacteria; Actinomycetes; Complementation; Mutation
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/BF00173728

We have characterized a mutant of Streptomyces clavuligerus NRRL 3585 which is almost completely blocked in cephalosporin biosynthesis and exhibits depressed activities of both the delta(L-alpha-aminoadipyl)-L-cysteinyl-D-valine (ACV) synthetase and cyclase enzymes of the cephalosporin pathway. A wild-type DNA region was cloned which partially restores antibiotic production, ACV synthetase and cyclase activities to this mutant. The recombinant plasmid exhibits a variable copy number in different transformants. Hybridization experiments indicate that sequences homologous to the cloned region are present in various beta-lactam-producing Streptomyces spp. but absent in species which are not known to produce this class of antibiotics. Furthermore, the chromosomal copy of the cloned region lies in close proximity to a gene coding for the isopenicillin N synthase gene of the cephalosphorin pathway.