Journal
ACS CHEMICAL BIOLOGY
Volume 11, Issue 7, Pages 1992-2001Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acschembio.6b00245
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Funding
- National Institutes of Health [AI065357 RM DP 008, AI089347]
- National Natural Science Foundation of China [31470787, 31170763]
- National Science Foundation [CHE-1429195]
- Division Of Chemistry
- Direct For Mathematical & Physical Scien [1429195] Funding Source: National Science Foundation
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The anthracycline natural product dutomycin and its precursor POK-MD1 were isolated from Streptomyces minoensis NRRL B-5482. The dutomycin biosynthetic gene cluster was identified by genome sequencing and disruption of the ketosynthase gene. Two polyketide synthase (PKS) systems are present in the gene cluster, including a type II PKS and a rare highly reducing iterative type I PKS. The type I PKS DutG repeatedly uses its active sites to create a nine-carbon triketide chain that is subsequently transferred to the alpha-L-axenose moiety of POK-MD1 at 4 ''-OH to yield dutomycin. Using a heterologous recombination approach, we disrupted a putative methyltransferase gene (dutMT1) and two glycosyltransferase genes (dutGT1 and dutGT2). Analysis of the metabolites of these mutants revealed the functions of these genes and yielded three dutomycin analogues SW140, SW91, and SW75. The major product SW91 in Streptomyces minoensis NRRL B-5482-Delta DutMT1 was identified as 12-desmethyl-dutomycin, suggesting that DutMT1 is the dedicated 12-methyltransferase. This was confirmed by the in vitro enzymatic assay. DutGT1 and DutGT2 were found to be responsible for the introduction of beta-D-amicetose and alpha-L-axenose, respectively. Dutomycin and SW91 showed strong antibacterial activity against Staphylococcus aureus and methicillin-resistant S. aureus, whereas POK-MD1 and SW75 had no obvious inhibition, which revealed the essential role of the C-4 '' triketide chain in antibacterial activity. The minimal inhibitory concentration of SW91 against the two strains was 0.125 mu g mL(-1), lower than that of dutomycin (0.25 mu g mL(-1)), indicating that the antibacterial activity of dutomycin can be improved through biosynthetic structural modification.
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