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Bioconversion of 12-, 14-, and 16-membered ring aglycones to glycosylated macrolides in an engineered strain of Streptomyces venezuelae

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To develop a system for combinatorial biosynthesis of glycosylated macrolides, Streptomyces venezuelae was genetically manipulated to be deficient in the production of its macrolide antibiotics by deletion of the entire biosynthetic gene cluster encoding the pikromycin polyketide synthases and desosamine biosynthetic enzymes. Two engineered deoxysugar biosynthetic pathways for the biosynthesis of thymidine diphosphate (TDP)-d-quinovose or TDP-d-olivose in conjunction with the glycosyltransferase–auxiliary protein pair DesVII/DesVIII derived from S. venezuelae were expressed in the mutant strain. Feeding the representative 12-, 14-, and 16-membered ring macrolactones including 10-deoxymethynolide, narbonolide, and tylactone, respectively, to each mutant strain capable of producing TDP-d-quinovose or TDP-d-olivose resulted in the successful production of the corresponding quinovose- and olivose-glycosylated macrolides. In mutant strains where the DesVII/DesVIII glycosyltransferase–auxiliary protein pair was replaced by TylMII/TylMIII derived from Streptomyces fradiae, quinovosyl and olivosyl tylactone were produced; however, neither glycosylated 10-deoxymethynolide nor narbonolide were generated, suggesting that the glycosyltransferase TylMII has more stringent substrate specificity toward its aglycones than DesVII. These results demonstrate successful generation of structurally diverse hybrid macrolides using a S. venezuelae in vivo system and provide further insight into the substrate flexibility of glycosyltransferases.

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We thank Professor Eric Cundliffe (University of Leicester) for kindly providing a tylactone standard. This work was supported by a grant (20050401-034-682-006-02-00) from the BioGreen 21 Program and grants from the National R&D Program for Cancer Control (0620300-1) and the SRC program of KOSEF through the Center for Intelligent Nano-Bio Materials at Ewha Womans University (grant R11-2005-008-00000-0). The authors wish to thank the Ministry of Education for the Brain Korea 21 fellowship.

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Correspondence to Yeo Joon Yoon.

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Won Seok Jung and Ah Reum Han contributed equally to this work.

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Jung, W.S., Han, A.R., Hong, J.S.J. et al. Bioconversion of 12-, 14-, and 16-membered ring aglycones to glycosylated macrolides in an engineered strain of Streptomyces venezuelae . Appl Microbiol Biotechnol 76, 1373–1381 (2007).

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  • Combinatorial biosynthesis
  • Streptomyces
  • Macrolide
  • Glycosyltransferase
  • Auxiliary protein