The ABC transporter Tba of Amycolatopsis balhimycina is required for efficient export of the glycopeptide antibiotic balhimycin.
All known gene clusters for glycopeptide antibiotic biosynthesis contain a conserved gene supposed to encode an ABC-transporter. In the balhimycin-producer Amycolatopsis balhimycina this gene (tba) is localised between the prephenate dehydrogenase gene pdh and the peptide synthetase gene bpsA. Inactivation of tba in A. balhimycina by gene replacement did not interfere with growth and did not affect balhimycin resistance. However, in the supernatant of the tba mutant RM43 less balhimycin was accumulated compared to the wild type; and the intra-cellular balhimycin concentration was ten times higher in the tba mutant RM43 than in the wild type. These data suggest that the ABC transporter encoded in the balhimycin biosynthesis gene cluster is not involved in resistance but is required for the efficient export of the antibiotic. To elucidate the activity of Tba it was heterologously expressed in Escherichia coli with an N-terminal His-tag and purified by nickel chromatography. A photometric assay revealed that His(6)-Tba solubilised in dodecylmaltoside possesses ATPase activity, characteristic for ABC-transporters.
Menges
R
R
Lehrstuhl für Mikrobiologie/Biotechnologie, Mikrobiologisches Institut, Universität Tübingen, Tübingen, Germany.
Muth
G
G
Wohlleben
W
W
Stegmann
E
E
D018528:ATP-Binding Cassette Transporters / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D000192:Actinomycetales / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000378:metabolism / Q000493:pharmacokinetics / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D001692:Biological Transport / Q000187:drug effects; D015139:Blotting, Southern; D024881:Drug Resistance, Bacterial; D004591:Electrophoresis, Polyacrylamide Gel; D006020:Glycopeptides / Q000378:metabolism / Q000493:pharmacokinetics / Q000494:pharmacology; D008969:Molecular Sequence Data; D009154:Mutation; D016133:Polymerase Chain Reaction; D017433:Protein Structure, Secondary; D014640:Vancomycin / Q000031:analogs & derivatives* / Q000378:metabolism / Q000493:pharmacokinetics / Q000494:pharmacology