CHARACTERIZATION OF STREPTOMYCES-NOGALATER GENES ENCODING ENZYMES INVOLVED IN GLYCOSYLATION STEPS IN NOGALAMYCIN BIOSYNTHESIS

The sno gene cluster in Streptomyces nogalater ATCC 27451 contains the nogalamycin biosynthesis genes. A set of plasmid constructions carryi ng fragments of the sno cluster that lie downstream of snoD were used to complement the S. galilaeus mutant H039, which is blocked in rhodos amine and 2-deoxyfucose biosynthesis in the aclacinomycin pathway. Seq uence analysis of this cluster revealed three contiguous open reading frames (ORFs) that were designated snoF, snoG, and snoH. Only those pl asmid constructs that expressed SnoG were able to complement H039. Sno G shows similarity to GalE, a UDP-glucose-4-epimerase catalyzing the e pimerization of UDP-glucose to UDP-galactose. The putative SnoF protei n is similar to 3,5-epimerases involved in rhamnose biosynthesis. The deduced product of snoH is a 489-amino acid polypeptide. It is similar to the product of dau ORF3 found in the daunomycin cluster. However i ts function is still unclear. Based on the complementation experiments and sequence analysis, this part of the size cluster is suggested to be involved in the biosynthesis of the sugar portion of nogalamycin. I nterestingly, SnoA, a transcriptional activator for the sno minimal po lyketide synthase, is also needed to express this cluster.