Open AccessIdentification of the Coumermycin A 1 Biosynthetic Gene Cluster of Streptomyces rishiriensis DSM 40489
Author(s) -
Zhaoxin Wang,
Shu-Ming Li,
Lutz Heide
Publication year - 2000
Publication title -
antimicrobial agents and chemotherapy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.07
H-Index - 259
eISSN - 1070-6283
pISSN - 0066-4804
DOI - 10.1128/aac.44.11.3040-3048.2000
Subject(s) - orfs , gene cluster , biology , genetics , novobiocin , cosmid , gene , dna gyrase , thiostrepton , open reading frame , escherichia coli , peptide sequence , ribosome , rna , antibiotics
The biosynthetic gene cluster of the aminocoumarin antibiotic coumermycin A1 was cloned by screening of a cosmid library ofStreptomyces rishiriensis DSM 40489 with heterologous probes from a dTDP-glucose 4,6-dehydratase gene, involved in deoxysugar biosynthesis, and from the aminocoumarin resistance gyrase genegyrB r . Sequence analysis of a 30.8-kb region upstream ofgyrB r revealed the presence of 28 complete open reading frames (ORFs). Fifteen of the identified ORFs showed, on average, 84% identity to corresponding ORFs in the biosynthetic gene cluster of novobiocin, another aminocoumarin antibiotic. Possible functions of 17 ORFs in the biosynthesis of coumermycin A1 could be assigned by comparison with sequences in GenBank. Experimental proof for the function of the identified gene cluster was provided by an insertional gene inactivation experiment, which resulted in an abolishment of coumermycin A1 production.