Affiliation:
1. Mikrobiologie/Biotechnologie1and
2. Institut für Organische Chemie,2 Eberhard-Karls-Universität Tübingen, D-72076 Tübingen, Germany
Abstract
ABSTRACT
Seven complete genes and one incomplete gene for the biosynthesis of the glycopeptide antibiotic balhimycin were isolated from the producer,
Amycolatopsis mediterranei
DSM5908, by a reverse-cloning approach and characterized. Using oligonucleotides derived from glycosyltransferase sequences, a 900-bp glycosyltransferase gene fragment was amplified and used to identify a DNA fragment of 9,882 bp. Of the identified open reading frames, three (
oxyA
to -
C
) showed significant sequence similarities to cytochrome P450 monooxygenases and one (
bhaA
) showed similarities to halogenase, and the genes
bgtfA
to -
C
showed similarities to glycosyltransferases. Glycopeptide biosynthetic mutants were created by gene inactivation experiments eliminating oxygenase and glycosyltransferase functions. Inactivation of the oxygenase gene(s) resulted in a balhimycin mutant (SP1-1) which was not able to synthesize an antibiotically active compound. Structural analysis by high-performance liquid chromatography–mass spectrometry, fragmentation studies, and amino acid analysis demonstrated that these oxygenases are involved in the coupling of the aromatic side chains of the unusual heptapeptide. Mutant strain HD1, created by inactivation of the glycosyltransferase gene
bgtfB
, produced at least four different compounds which were not glycosylated but still antibiotically active.
Publisher
American Society for Microbiology
Subject
Infectious Diseases,Pharmacology (medical),Pharmacology
Cited by
205 articles.
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