A two-plasmid system for the glycosylation of polyketide antibiotics: bioconversion of ε-rhodomycinone to rhodomycin D

Abstract

The high-yielding glycosylation of the aromatic polyketide epsilon-rhodomycinone using plasmid-borne deoxysugar biosynthesis genes proves that the minimal information for L-daunosamine biosynthesis and attachment in the heterologous host is encoded by the dnmLMJVUTS genes. This is a general approach to making both known and new glycosides of anthracyclines, several of which have medically important antitumor activity.

Biosynthesis of the dideoxysugar component of jadomycin B: genes in the jad cluster of Streptomyces venezuelae ISP5230 for l-digitoxose assembly and transfer to the angucycline aglycone The GenBank accession number for the sequence reported in this paper is AY026363.

“…As in the present investigation, knowledge of biosynthetic reactions is limited to prediction of enzyme functions based on gene sequences, their relationship to the existing database and the phenotypic changes that result from specific gene disruptions. Biochemical characterization of intermediates accumulated after gene inactivation provides supporting evidence for enzyme function and has not only cast light on several sugar biosynthetic pathways, but also helped in the design of hybrid natural compounds (Zhao et al, 1998 ;Kunzel et al, 1999 ;Olano et al, 1999).…”

“…Novel metabolites can be generated by interchanging secondary metabolic biosynthesis genes among a variety of micro-organisms, or by creating hybrid genes that direct the synthesis of unique and hitherto unavailable structures. Since Hopwood & Sherman (1990) reviewed polyketide antibiotic synthesis with a focus on PKS genes, there has been substantial progress in recombining genes to construct novel polyketide aglycones (Hutchinson 1999 ;McDaniel et al, 1999 ;David et al, 1998 ;Shen et al, 1999) ; recent extensions of this approach to include deoxysugar biosynthesis genes expand opportunities to genetically engineer new microbial metabolites with medical applications (Olano et al, 1999 ;Gaisser et al, 2000).…”

Natural‐Product Sugar Biosynthesis and Enzymatic Glycodiversification

“…The encoded enzymes likely catalyze the corresponding steps in both sugar biosynthetic pathways. [62] The sugars l-nogalamine (52), [22] l-daunosamine (see 53), [76] and l-rhodosamine (see 54) [77] are found in the anthracycline antibiotics nogalamycin, daunorubicin, and aclarubicin, respectively. Their common precursor is 60, which undergoes 3,5-epimerization and stereospecific ketoreduction in each pathway.…”

“…Interestingly, l-oleandrose is constructed by different routes in these two pathways. It was shown by heterologous expression of the oleandomycin biosynthetic genes [97] that 77 is formed from 73 by 3,5-epimerization and 4-ketoreduction catalyzed by OleL and OleU, respectively, resulting in TDP-l-olivose (76), which is the donor for glycosyltransfer. 3-O-Methylation by OleY has been confirmed in vitro to occur after sugar attachment.…”

Combinatorial Biosynthesis of Antitumor Deoxysugar Pathways in Streptomyces griseus : Reconstitution of “Unnatural Natural Gene Clusters” for the Biosynthesis of Four 2,6- d -Dideoxyhexoses

“…This type of modification can be approached by combinatorial biosynthesis, providing that flexible glycosyltransferases exist, together with the possibility of using differ-ent nucleotide DOHs. In this context, the isolation of natural gene clusters and the reconstitution of "unnatural gene clusters" for the biosynthesis of DOHs have allowed their use for the generation of new glycosylated compounds (23,26,38). In this study, we have reconstituted "unnatural natural gene clusters" for the biosynthesis of four different activated 2,6-DDOHs, namely, TDP-D-olivose, TDP-D-oliose, TDP-D-digitoxose, and TDP-D-boivinose.…”