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Volume 147, Issue 9

The VanY DD-carboxypeptidase of BM4339 is a penicillin-binding protein Free

Abstract

VanD-type BM4339 is constitutively resistant to vancomycin and to low levels of teicoplanin. This strain produces peptidoglycan precursors terminating in D-lactate but, unlike VanA- and VanB-type strains, BM4339 has a mutated ligase gene and cannot synthesize D-Ala-D-Ala. Consequently, although it possesses and genes, it should not require an active VanX-type DD-dipeptidase or a VanY-type DD-carboxypeptidase for resistance. The gene contains the signatures of a penicillin-binding protein (PBP) and is believed to encode a penicillin-sensitive DD-carboxypeptidase. The enzyme activity was found to be membrane-bound and inhibited by low concentrations of benzylpenicillin in membrane preparations and in intact bacteria, indicating that the active site was present on the outside surface of the membrane. The 38 kDa protein was revealed as a PBP present in more copies per cell than conventional PBPs and all the protein was accessible to benzylpenicillin added externally, confirming the localization of the active site. A glycopeptide-susceptible strain of lacked this PBP, and the membrane-bound DD-carboxypeptidase activity was less than 5% of that of BM4339. Although the active site of VanY was external to the membrane, UDP-MurNAc-tetrapeptide was produced internally, probably from UDP-MurNAc-pentadepsipeptide. The presence of benzylpenicillin at low concentrations in the growth medium substantially reduced the amount of tetrapeptide produced, indicating that inhibition of VanY by benzylpenicillin influenced production of peptidoglycan precursors internally. A model to explain these contrasting observations is proposed.

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Keyword(s): glycopeptide resistance , membrane-bound dd-peptidase , PBP, penicillin-binding protein , vancomycin resistance and VanD phenotype
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2001-09-01
2024-04-18
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The VanYD DD-carboxypeptidase of Enterococcus faecium BM4339 is a penicillin-binding protein
Microbiology 147, 2571 (2001); https://doi.org/10.1099/00221287-147-9-2571
/content/journal/micro/10.1099/00221287-147-9-2571
/content/journal/micro/10.1099/00221287-147-9-2571
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