Isolation and characterization of bluensomycin biosynthetic genes from Streptomyces bluensis.
The biosynthetic gene cluster for bluensomycin, a member of the aminoglycoside family of antibiotics, was isolated and characterized from the bluensomycin producing strain, Streptomyces bluensis ATCC27420. PCR primers were designed specifically to amplify a segment of the dTDP-glucose synthase gene based on its conserved sequences among several actinomycete strains. By screening a cosmid library using amplified PCR fragments, a 30-kb DNA fragment was isolated. Sequence analysis identified 15 open reading frames (ORFs), eight of which had previously been identified by Piepersberg et al. But seven are novel to this study. We demonstrated that one of these ORFs, blmA, confers resistance against the antibiotic dihydrostreptomycin, and another, blmD, encodes a dTDP-glucose synthase. These findings suggest that the isolated gene cluster is very likely to be responsible for the biosynthesis of bluensomycin.
Jung
Yong Gyun
YG
Department of Biological Science, Institute of Bioscience and Biotechnology, Myong Ji University, 449-728, Yongin, South Korea/
Kang
Suk Ho
SH
Hyun
Chang Gu
CG
Yang
Young Yell
YY
Kang
Choong Min
CM
Suh
Joo Won
JW
D002240:Carbohydrate Sequence; D003001:Cloning, Molecular; D004096:Dihydrostreptomycin Sulfate / Q000031:analogs & derivatives* / Q000276:immunology / Q000378:metabolism*; D005798:Genes, Bacterial; D005947:Glucose / Q000031:analogs & derivatives* / Q000378:metabolism; D005958:Glucosephosphates / Q000378:metabolism; D008359:Mannose-6-Phosphate Isomerase / Q000032:analysis; D005810:Multigene Family; D016254:Mutagenesis, Insertional; D009713:Nucleotidyltransferases / Q000032:analysis; D016366:Open Reading Frames; D013302:Streptomyces / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D013942:Thymine Nucleotides / Q000378:metabolism
D005958:Glucosephosphates; D013942:Thymine Nucleotides; C103522:deoxythymidine diphosphate-glucose; C031590:glucose-1-phosphate; D009713:Nucleotidyltransferases; C085243:dTDP-D-glucose synthase; D008359:Mannose-6-Phosphate Isomerase; D005947:Glucose; C006786:bluensomycin; D004096:Dihydrostreptomycin Sulfate