FIELD: biotechnology, biochemistry, molecular biology, genetic engineering.
SUBSTANCE: invention relates to preparing new polyketide synthase that is necessary for biosynthesis of epotilones A and B. Invention describes the isolated nucleic acid encoding domain of β-ketoacyl synthase involves to biosynthesis of epotilones and its variants. Nucleic acid is isolated, in particular, from Sorangium cellulosum. Recombinant vector is constructed comprising chimeric gene with heterologous promoter sequence associated functionally with isolated molecule of nucleic acid. Method for expression of epotilone is carried out using recombinant host, for example, microorganism E. coli, clone pEP014, deposited at № NRRL B-30033 in the process of its culturing under suitable conditions. Also, invention provides preparing epotilone by extraction from recombinant host. Isolated polypeptide involves to biosynthesis of epotilones comprises domain of β-ketoacyl synthase, acyltransferase, enoyl-reductase, acyl-protein carrier, dehydratase, β-keto-reductase, methyltransferase, thioesterase or non-ribisomal acid-amino acid-ligase (NRPSs). Invention provides realization of preparing epotilones in the industrial scale that elicit antifungal activity and high cytotoxicity with respect to mammalian cell culture, among them, against tumor cells.
EFFECT: improved preparing method, valuable medicinal properties of epotilones.
61 cl, 4 tbl, 15 ex
