Mycolic acid biosynthesis and enzymic characterisation of the β-ketoacyl-ACP synthase A-condensing enzyme from Mycobacterium tuberculosis

L Kremer, Lynn Dover, S Carrere, KM Nampoothiri, S Lesjean, AK Brown, PJ Brennan, David Minnikin, C Locht, Gurdyal Besra

Research output: Contribution to journalArticle

97 Citations (Scopus)

Abstract

Mycolic acids consist of long-chain alpha-alkyl-beta-hydroxy fatty acids that are produced by successive rounds of elongation catalysed by a type II fatty acid synthase (FAS-II). A key feature in the elongation process is the condensation of a two-carbon unit from malonyl-acyl-carrier protein (ACP) to a growing acyl-ACP chain catalysed by a beta-ketoacyl-ACP synthase (Kas). In the present study, we provide evidence that kasA from Mycobacterium tuberculosis encodes an enzyme that elongates in vivo the meromycolate chain, in both Mycobacterium smegmatis and Mycobacterium chelonae. We demonstrate that KasA belongs to the FAS-II system, which utilizes primarily palmitoyl-ACP rather than short-chain acyl-ACP primers. Furthermore, in an in vitro condensing assay using purified recombinant KasA, palmitoyl-AcpM and malonyl-AcpM, KasA was found to express Kas activity. Also, mutated KasA proteins, with mutation of Cys(171), His(311), Lys(340) and His(345) to Ala abrogated the condensation activity of KasA in vitro completely. Finally, purified KasA was highly sensitive to cerulenin, a well-known inhibitor of Kas, which may lead to the development of novel anti-mycobacterial drugs targeting KasA.
Original languageEnglish
Pages (from-to)423-430
Number of pages8
JournalBiochemical Journal
Volume364
Issue number2
DOIs
Publication statusPublished - 1 Jun 2002

Keywords

  • acyl-carrier protein
  • fatty acid
  • cerulenin
  • type II fatty acid synthase

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