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Genetics and Molecular Biology |
Department of Biology, Dalhousie University, Halifax, Nova Scotia, Canada B3H 4J11
School of Pharmacy, University of Wisconsin, Madison, WI53706, USA2
Author for correspondence: L. C. Vining. Tel: +1 902 494 2040. Fax: +1 902 494 3736. e-mail: Leo.Vining{at}Dal.Ca
Analysis of a region of chromosomal DNA lying between jadR1 and jadI in the gene cluster for jadomycin biosynthesis in Streptomyces venezuelae ISP5230 detected an ORF encoding 584 amino acids similar in sequence to the biotin carboxylase (BC) and biotin carboxyl carrier protein (BCCP) components of acyl-coenzyme A carboxylases. Multiple sequence alignments of the deduced Jad protein with acyl-coenzyme A carboxylases from various sources located the BC and BCCP components in the N- and C-terminal regions, respectively, of the deduced polypeptides. The organization and amino acid sequence of the deduced polypeptide most closely resembled those in other Gram-positive bacteria broadly classified as actinomycetes. Disrupting the gene, designated jadJ, severely reduced but did not eliminate jadomycin production. The disruption had no effect on growth or morphology of the organism, implying that the product of jadJ is not essential for fatty acid biosynthesis. It is concluded that jadJ supplies malonyl-coenzyme A for biosynthesis of the polyketide intermediate that is eventually processed to form the antibiotic jadomycin B.
Keywords: acetyl-coenzyme A carboxylase genes, jadomycin biosynthesis, Streptomyces venezuelae
Abbreviations: ACC, acyl-coenzyme A carboxylase; BC, biotin carboxylase; BCCP, biotin carboxyl carrier protein; DIG, digoxigenin; PKS, polyketide synthase; TC, transcarboxylase
The GenBank accession number for the sequence reported in this paper is AF126429.
a Present address: 306/C1-295 Dow AgroSciences LLC, 9330 Zionsville Road, Indianapolis, IN 46268-1054, USA.
b Present address: Department of Microbiology, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z3.
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