Cloning and characterization of a glycosyltransferase gene involved in the biosynthesis of anthracycline antibiotic β-rhodomycin from Streptomyces violaceus

Abstract

A glycosyltransferase gene, rhoG, involved in the biosynthesis of the anthracycline antibiotic L-rhodomycin was isolated as a 4.1-kb DNA fragment containing rhoG and its flanking region from Streptomyces violaceus by degenerate and inverse PCR. Sequencing analysis showed that rhoG was located in a gene cluster involved in the biosynthesis of the constitutive deoxysugar of L-rhodomycin. The function of rhoG was verified by gene disruption, which was generated by replacing the internal 0.9-kb region of S. violaceus chromosome with a fragment including the SacI-blunted region. The rhoG disruption resulted in complete loss of L-rhodomycin productivity, along with the accumulation of a non-glycosyl intermediate O-rhodomycinone. In addition, the complementation test demonstrated that rhoG restored L-rhodomycin production in this gene disruptant. These results indicated that rhoG is the glycosyltransferase gene responsible for the glycosylation of O-rhodomycinone in L-rhodomycin biosynthesis. ß

Insights in the glycosylation steps during biosynthesis of the antitumor anthracycline cosmomycin: characterization of two glycosyltransferase genes

“…CosG shows similarities with several glycosyltransferases which transfer aminodeoxysugars like RhoG from the rhodomycin pathway in S. violaceus (Miyamoto et al 2002), DnrS from the daunorubicin pathway in S. peucetius (Otten et al 1995), AclS from the aclacinomycin pathway in S. galilaeus (Chung et al 2002), and DesVII from the pikromycin pathway in S. venezuelae (Xue et al 1998). Based on this, CosG is the glycosyltransferase responsible for L-rhososamine transfer to the 7-or 10-position (or both) in the cosmomycin aglycone.…”

“…1) are an interesting group of compounds because they show one of the most complex glycosylation patterns found in anthracyclines with two trisaccharide chains attached at 7-and 10-positions (Miyamoto et al 2002). These compounds are produced by Streptomyces olindensis originally described as a producer of an anthracycline complex called retamycin (Gonçalves de Lima et al 1969).…”

Cloning, expression and functional analysis of nicotinate dehydrogenase gene cluster from Comamonas testosteroni JA1 that can hydroxylate 3-cyanopyridine

“…Alignment analysis showed the sequence contained the stop codon. Finally, the entire sequence encoding NaDH JA1 was amplified with primers JA-SLM-S and JA-SLM-F, cloned with TA cloning vector pMD18-TndhSLM, and sequenced (Miyamoto et al 2002).…”

Caracterização dos mecanismos de resistência ao antibiótico antitumoral cosmomicina D.

“…Estudos estruturais determinaram que a molécula é um complexo aromático com duas cadeias trissacarídicas no carbono 7 e 10 chamado cosmomicina D (Figura 8 ) (GARRIDO et al, 2006;FURLAN et al, 2004;). As cosmomicinas são um grupo de compostos interessantes porque apresentam um dos padrões de glicosilação mais complexos encontrados nas antraciclinas (MIYAMOTO et al, 2002).…”