Abstract
Actinomycetes produce many bioactive compounds with clinical, veterinary, or agricultural applications. Many of these natural products contain sugars attached to the corresponding agly-cons, which usually participate in the molecular recognition of the cellular target. The glycosy-lation pattern of these complex metabolites can be altered by chemical synthesis or synthetic modification of intermediates usually produced via fermentation and by the use of genetically engineered recombinant microorganisms and biosynthetic genes for in vivo experiments through combinatorial biosynthesis. Here, we describe procedures to generate novel glycosylated compounds derived from the antitumor compound elloramycin. This procedure involves biotransfor-mation of different recombinant strains of Streptomyces albus harboring the elloramycin ElmGT glycosyltransferase and able to synthesize different dTDP-activated deoxysugars whose biosynthesis is directed by different plasmids.
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References
Méndez, C. and Salas, J. A. (2001) Altering the glycosylation pattern of bioactive compounds. Trends Biotechnol. 19, 449–456.
Decker, H., Rohr, J., Motamedi, H., Zahner, H., and Hutchinson, C. R. (1995) Identification of Streptomyces olivaceus Tu 2353 genes involved in the production of the polyketide elloramycin. Gene 166, 121–126.
Blanco, G., Patallo, E. P., Braña, A. E, et al. (2001) Identification of a sugar flexible glycosyltransferase from Streptomyces olivaceus, the producer of the antitumor polyketide elloramycin. Chem. Biol. 8, 253–263.
Patallo, E. P., Blanco, G., Fischer, C., et al (2001) Deoxysugar methylation during biosynthesis of the antitumor polyketide elloramycin by Streptomyces olivaceus: characterization of three methyltransferase genes. J. Biol. Chem. 276, 18,765–18,774.
Aguirrezabalaga, I., Olano, C., Allende, N., et al. (2000) Identification and expression of genes involved in biosynthesis of L-oleandrose and its intermediate L-olivose in the oleandomycin producer Streptomyces antibioticus. Antimicrob. Agents Chemother. 44, 1266–1275.
Kuhstoss, S., Richardson, M. A., and Rao, R. N. (1991) Plasmid cloning vectors that integrate site-specifically in Streptomyces spp. Gene 97, 143–146.
Quiros, L. M., Aguirrezabalaga, I., Olano, C, Mendez, C., and Salas, J. A. (1998) Two glycosyltransferases and a glycosidase are involved in oleandomycin modification during its biosynthesis by Streptomyces antibioticus. Mol. Microbiol. 28, 1177–1185.
RodrÍguez, L., Oelkers, C., Aguirrezabalaga,., et al. (2000) Generation of hybrid elloramycin analogs by combinatorial biosynthesis using genes from anthracy-cline-type and macrolide biosynthetic pathways. J. Mol. Microbiol. Biotechnol. 2, 271–276.
Sambrook, J., Fritsch, E. E, and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
Kieser, T., Bibb, M. J., Buttner, M. J., Chater, K. E, and Hopwood, D. A. (2000) Practical Streptomyces genetics. The John Innes Foundation, Norwich, UK.
Vara, J., Lewandowska-Skarbek, M., Wang, Y. G., Donadio, S., and Hutchinson, C. R. (1989) Cloning of genes governing the deoxysugar portion of the erythromycin biosynthesis pathway in Saccharopolyspora erythraea (Streptomyces erythreus). J. Bacteriol. 171, 5872–5881.
Janssen, G. R. and Bibb, M. J. (1993) Derivatives of pUC18 that have BglII sites flanking a modified multiple cloning site and that retain the ability to identify recombinant clones by visual screening of Escherichia coli colonies. Gene 124, 133–134.
RodrÍguez, L., RodrÍguez, D., Olano, C., Braña, A. E, Mendez, C., and Salas, J. A. (2001) Functional analysis of OleY L-oleandrosyl 3-O-methyltransferase of the oleandomycin biosynthetic pathway in Streptomyces antibioticus. J. Bacteriol. 183, 5358–5363.
Vieira, J. and Messing, J. (1991) New pUC-derived cloning vectors with different selectable markers and DNA replication origins. Gene 100, 189–194.
RodrÍguez, L., Aguirrezabalaga, I., Allende, N., Braña, A. E, Mendez, C., and Salas, J. A. (2002) Engineering deoxysugar biosynthetic pathways from antibiotic-producing microorganisms. A tool to produce novel glycosylated bioactive compounds. Chem. Biol. 9, 721–729.
Wohlert, S.-E., Blancò, G., Lombo, E, et al. (1998) Novel hybrid tetracenomycins through combinatorial biosynthesis using a glycosyltransferase encoded by the elm-genes in cosmid 16F4 which shows a broad sugar substrate specificity. J. Am. Chem. Soc. 41, 10,596–10,601.
Fernandez, E., Weissbach, U., Sanchez Reillo, C., et al. (1998) Identification of two genes from Streptomyces argillaceus encoding glycosyltransferases involved in transfer of a disaccharide during biosynthesis of the antitumor drug mithramycin. J. Bacteriol. 180, 4929–4937.
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Méndez, C., Salas, J.A. (2005). Recombinant Microorganisms for the Biosynthesis of Glycosylated Antitumor Compounds. In: Barredo, JL. (eds) Microbial Processes and Products. Methods in Biotechnology, vol 18. Humana Press. https://doi.org/10.1385/1-59259-847-1:131
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DOI: https://doi.org/10.1385/1-59259-847-1:131
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