Characterization and complementation of a cephalosporin-deficient mutant of Streptomyces clavuligerus NRRL 3585.
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.
Piret
J
J
Department of Biology, Northeastern University, Boston, MA 02115.
Resendiz
B
B
Mahro
B
B
Zhang
J Y
JY
Serpe
E
E
Romero
J
J
Connors
N
N
Demain
A L
AL
D001426:Bacterial Proteins / Q000378:metabolism; D002511:Cephalosporins / Q000096:biosynthesis*; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics; D005816:Genetic Complementation Test; D009154:Mutation; D010088:Oxidoreductases / Q000235:genetics; D010453:Peptide Synthases / Q000235:genetics; D010641:Phenotype; D010957:Plasmids / Q000235:genetics; D015183:Restriction Mapping; D013302:Streptomyces / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D014169:Transformation, Bacterial
D001426:Bacterial Proteins; D002511:Cephalosporins; D004269:DNA, Bacterial; D010088:Oxidoreductases; C038370:isopenicillin N synthetase; D010453:Peptide Synthases; C095368:alpha-aminoadipyl-cysteinyl-valine synthetase