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Journal of Bacteriology, December 1999, p. 7161-7167, Vol. 181, No. 23
Department of Molecular Microbiology,
Washington University School of Medicine, St. Louis, Missouri
63110,1 and Department of Microbiology,
Albert Szent-Gyorgyi Medical University, Szeged POB-8-6701,
Hungary2
Received 20 May 1999/Accepted 10 September 1999
Analysis by two-dimensional gel electrophoresis revealed that
Mycobacterium avium expresses several proteins unique to an intracellular infection. One abundant protein with an apparent molecular mass of 50 kDa was isolated, and the N-terminal sequence was
determined. It matches a sequence in the M. tuberculosis
database (Sanger) with similarity to the enzyme isocitrate lyase of
both Corynebacterium glutamicum and Rhodococcus
fascians. Only marginal similarity was observed between this open
reading frame (ORF) (termed icl) and a second distinct ORF
(named aceA) which exhibits a low similarity to other
isocitrate lyases. Both ORFs can be found as distinct genes in the
various mycobacterial databases recently published. Isocitrate lyase is
a key enzyme in the glyoxylate cycle and is essential as an
anapleurotic enzyme for growth on acetate and certain fatty acids as
carbon source. In this study we express and purify Icl, as well as AceA
proteins, and show that both exhibit isocitrate lyase activity. Various
known inhibitors for isocitrate lyase were effective. Furthermore, we
present evidence that in both M. avium and M. tuberculosis the production and activity of the isocitrate lyase
is enhanced under minimal growth conditions when supplemented with
acetate or palmitate.
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Characterization of Activity and Expression of
Isocitrate Lyase in Mycobacterium avium and
Mycobacterium tuberculosis
*
Corresponding author. Mailing address: Department of
Molecular Microbiology, Washington University Medical School, 660 S. Euclid Ave., Box 8230, St. Louis, MO 63110. Phone: (314) 362-4987. Fax:
(314) 362-1232. E-mail: hoenerk{at}borcim.wustl.edu.
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