Cloning of modular type I polyketide synthase genes from salinomycin producing strain of Streptomyces albus.
Cloning of polyether polyketide synthase (PKS) genes for salinomycin biosynthesis was attempted from Streptomyces albus. Seven beta-ketoacyl synthase (KS) core regions were obtained by PCR amplification using primers designed based on the conserved KS domains of type I PKSs. Using the KS fragment as a probe, screening of an S. albus genomic DNA library was carried out by colony hybridization. From the positive cosmid clone isolated, a 4.5-kbBamHI fragment was subcloned and sequenced. It showed high homology with bacterial type I PKSs and was deduced to code for KS, malonyl transferase, and ketoreductase motifs. By gene disruption with this 4.5-kb BamHI fragment, the cloned gene was shown to be a part of the salinomycin biosynthetic gene cluster of S. albus.
Izumikawa
Miho
M
Department of Chemistry, School of Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, 113-0033, Tokyo, Japan.
Murata
Michio
M
Tachibana
Kazuo
K
Ebizuka
Yutaka
Y
Fujii
Isao
I
D000595:Amino Acid Sequence; D001665:Binding Sites; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D015698:Genomic Library; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000737:chemistry / Q000235:genetics*; D009154:Mutation / Q000235:genetics; D017434:Protein Structure, Tertiary; D011714:Pyrans / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D013302:Streptomyces / Q000145:classification / Q000201:enzymology* / Q000235:genetics*; D014169:Transformation, Bacterial; D000095662:Polyether Polyketides
D009097:Multienzyme Complexes; D011714:Pyrans; C010327:salinomycin; D000095662:Polyether Polyketides