Biosynthetic Gene Cluster of the Glycopeptide Antibiotic Teicoplanin

Abstract

The gene cluster encoding biosynthesis of the clinically important glycopeptide antibiotic teicoplanin has been cloned from Actinoplanes teichomyceticus. Forty-nine putative open reading frames (ORFs) were identified within an 89 kbp genetic locus and assigned roles in teicoplanin biosynthesis, export, resistance, and regulation. Two ORFs, designated orfs 1 and 10*, showed significant homology to known glycosyltransferases. When heterologously expressed in Escherichia coli, these glycosyltransferases were shown to catalyze the transfer of UDP-(N-acetyl)-glucosamine onto, respectively, 3-chloro-beta-hydroxytyrosine-6 (3-Cl-6betaHty) and 4-hydroxyphenylglycine-4 (4Hpg) of the teicoplanin heptapeptide aglycone. The product of another ORF, orf11*, was demonstrated in vitro to transfer n-acetyl-, n-butyryl-, and n-octanoyl-groups from acyl-CoA donors either to a free UDP-aminosugar or to an aminosugar moiety in the teicoplanin pseudoaglycone, thus identifying Orf11* as the key acyltransferase in teicoplanin maturation. These findings should accelerate the combinatorial engineering of new and improved glycopeptide drugs.

Characterization of a regiospecific epivancosaminyl transferase GtfA and enzymatic reconstitution of the antibiotic chloroeremomycin

“…During balhimycin biosynthesis, bGtfA transfers the oxovancosaminyl moiety after OH-PheGly 4 has been glucosylated by bGtfB, because the bGtfB-deficient mutant accumulates only heptapeptide intermediate (30). In contrast, the glycosyltransferases (tGtfA and tGtfB) from a teicoplaininproducing Actinoplanes teichomyceticus ATCC31131 displayed no preferential glycosylation sequence in decorating the aglycone backbone (31,32). The glycosylation sequence of chloroeremomycin is also supported by the crystal structures of GtfA and GtfD.…”

Oxazolomycin Biosynthesis in Streptomyces albus JA3453 Featuring an “Acyltransferase-less” Type I Polyketide Synthase That Incorporates Two Distinct Extender Units

“…OzmR shows significant similarity to the LysR family of regulators, such as Orf5 (accession number BAA32133; 53% identity) from Streptomyces griseus (38), whereas OzmU belongs to the SARP family of regulators, including Orf31* (accession number CAG15043; 37% identity) involved in glycopeptide teicoplanin biosynthesis in Actinoplanes teichomyceticus (39), AfsR (accession number BAA14186; 33% identity) in Streptomyces coelicolor, a pleiotropic antibiotic regulator (40), and RubS (accession number AAM97369; 30% identity) involved in rubromycin biosynthesis in Streptomyces collinus. Although OzmT resembles other Thr-tRNA synthetases, such as SCH63.25 (CAC 10316, 77/83) in S. coelicolor, its exact roles still need to be determined.…”

A gene cluster for the biosynthesis of moenomycin family antibiotics in the genome of teicoplanin producer Actinoplanes teichomyceticus

“…Teichomycin A1 (TeiA) complex was shown to fall into moenomycin family of antibiotics (Bardone et al 1978;Borghi et al 1984), while teichomycin A2 turned out to be a mixture of lipoglycopeptide metabolites known today as teicoplanin (Pryka et al 1988;Li et al 2004;Sosio et al 2004;Horbal et al 2014). From limited chemical degradation experiments (Bardone et al 1978;Borghi et al 1984) it was inferred that TeiA closely resembles other members of moenomycin family, although the exact structure of TeiA was not established.…”