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.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Bierman M, Logan R, O’Brien K, Seno ET, Rao RN, Schoner BE (1992) Plasmid cloning vectors for the conjugal transfer of DNA from Escherichia coli to Streptomyces spp. Gene 116:43–49
Blanchard S, Thorson JS (2006) Enzymatic tools for engineering natural product glycosylation. Curr Opin Chem Biol 10:263–271
Borisova SA, Zhao L, Sherman DH, Liu HW (1999) Biosynthesis of desosamine: construction of a new macrolide carrying a genetically designed sugar moiety. Org Lett 1:133–136
Borisova SA, Zhao L, Melançon CE, Kao CL, Liu HW (2004) Characterization of the glycosyltransferase activity of DesVII: analysis and implication for the biosynthesis of macrolide antibiotics. J Am Chem Soc 126:6534–6535
Borisova SA, Zhang C, Takahashi H, Zhang H, Wong AW, Thorson JS, Liu HW (2006) Substrate specificity of the macrolide-glycosylating enzyme pair DesVII/DesVIII: opportunity, limitations, and mechanistic hypotheses. Angew Chem Int Ed 45:2748–2753
Gaisser S, Reather J, Wirtz G, Kellenberger L, Staunton J, Leadlay P (2000) A defined system for hybrid macrolide biosynthesis in Saccharopolyspora erythraea. Mol Microbiol 36:391–401
Hong JSJ, Park SH, Choi CY, Sohng JK, Yoon YJ (2004) New olivosyl derivatives of methymycin/pikromycin from an engineered strain of Streptomyces venezuelae. FEMS Microbiol Lett 238:291–399
Hong JSJ, Kim WS, Lee SK, Koh HS, Park HS, Park SJ, Kim YS, Yoon YJ (2005) The role of a second protein (DesVIII) in glycosylation for the biosynthesis of hybrid macrolide antibiotics in Streptomyces venezuelae. J Microbiol Biotechnol 15:640–645
Hong JSJ, Park SJ, Parajuli N, Park SR, Koh HS, Jung WS, Choi CY, Yoon YJ (2007) Functional analysis of DesVIII homologues involved in glycosylation of macrolide antibiotics by interspecies complementation. Gene 386:123–130
Jung WS, Lee SK, Hong JSJ, Rark SR, Jeong SJ, Han AR, Sohng JK, Kim BG, Choi CY, Sherman DH, Yoon YJ (2006) Heterologous expression of tylosin polyketide synthase and production of a hybrid macrolide in Streptomyces venezuelae. Appl Microbiol Biotechnol 72:763–769
Kieser T, Bibb MJ, Buttner MJ, Chater KF, Hoopwood DA (2000) Practical Streptomyces genetics. John Innes Centre, Norwich
Lee SK, Park JW, Kim JW, Jung WS, Park SR, Choi CY, Kim ES, Kim BS, Ahn JS, Sherman DH, Yoon YJ (2006) Neopikromycin and novapikromycin from the pikromycin biosynthetic pathway of Streptomyces venezuelae. J Nat Prod 69:847–849
Melançon CE, Liu HW (2007) Engineered biosynthesis of macrolide derivatives bearing the non-natural deoxysugars 4-epi-D-mycaminose and 3-N-monomethylamino-3-deoxy-D-fucose. J Am Chem Soc 129:4896–4897
Melançon CE, Takahashi H, Liu HW (2004) Characterization of tylM3/tylM2 and mydC/mycB pairs required for efficient glycosyltransfer in mcrolide antibiotic biosynthesis. J Am Chem Soc 126:16725–16727
Melançon CE, Yu WL, Liu HW (2005) TDP-mycaminose biosynthetic pathway revised and conversion of desosamine pathway to mycaminose pathway with one gene. J Am Chem Soc 127:12240–12241
Méndez C, Salas JA (2001) Altering the glycosylation pattern of bioactive compounds. Trends Biotechnol 11:449–456
Pérez M, Lombó F, Baig I, Braña AF, Rohr J, Salas JA, Méndez C (2006) Combinatorial biosynthesis of antitumor deoxysugar pathways in Streptomyces griseus: reconstitution of “unnatural natural gene cluster” for the biosynthesis of four 2,6-D-dideoxyhexoses. Appl Environ Microbiol 72:6644–6652
Reeves CD, Ward SL, Revill WP, Suzuki H, Marcus M, petrakovsky OV, Marquez S, Fu H, Dong SD, Katz L (2004) Production of hybrid 16-membered macrolides by expressing combinations of polyketide synthease genes in engineered Streptomyces fradiae hosts. Chem Biol 11:1465–1472
Rodriguez E, McDaniel R (2001) Combinatorial biosynthesis of antimicrobials and other natural products. Cur Opin Microbiol 4:526–534
Sambrook J, Fritsch EF, Maniatis T (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY
Tang L, McDaniel R (2001) Construction of desosamine containing polyketide libraries using a glycosyltransferase with broad substrate specificity. Chem Biol 8:547–555
Xue Y, Sherman DH (2001) Biosynthesis and combinatorial biosynthesis of pikromycin-related macrolides in Streptomyces venezuelae. Metab Eng 3:15–26
Yamase H, Zhao L, Liu HW (2000) Engineering a hybrid sugar biosynthetic pathway: production of L-rhamnose and its implication on dihydrostreptose biosynthesis. J Am Chem Soc 122:12397–12398
Yoon YJ, Beck JB, Kim BS, Kang HY, Reynolds KA, Sherman DH (2002) Generation of multiple bioactive macrolides by hybrid modular polyketide synthases in Streptomyces venezuelae. Chem Biol 9:203–214
Zhao L, Sherman DH, Liu HW (1998a) Biosynthesis of desosamine: construction of a new methymycin/neomethymycin analogue by deletion of a desosamine biosynthetic gene. J Am Chem Soc 120:10256–10257
Zhao L, Que NLS, Xue Y, Sherman DH, Liu HW (1998b) Mechanistic studies of desosamine biosynthesis: C-4 deoxygenation precedes C-3 transamination. J Am Chem Soc 120:12159–12160
Zhao L, Borisova SA, Yeung SM, Liu HW (2001) Study of C-4 deoxygenation in the biosynthesis of desosamine: evidence implicating a novel mechanism. J Am Chem Soc 123:7909–7910
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.
Won Seok Jung and Ah Reum Han contributed equally to this work.
About this article
Cite this article
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). https://doi.org/10.1007/s00253-007-1101-y
- Combinatorial biosynthesis
- Auxiliary protein