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Applied and Environmental Microbiology, July 2000, p. 2981-2987, Vol. 66, No. 7
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Cloning of the Malic Enzyme Gene from Corynebacterium glutamicum and Role of the Enzyme in Lactate Metabolism

Pierre Gourdon,1 Marie-France Baucher,2 Nic D. Lindley,1,* and Armel Guyonvarch2

Laboratoire de Biotechnologie-Bioprocédés, UMR INSA/CNRS 5504 and UMR INRA 792, Centre de Bioingénierie Gilbert Durand, Institut National des Sciences Appliqueés, 31077 Toulouse Cedex,1 and Institut de Génétique et Microbiologie, UMR CNRS 8621, Université Paris-Sud, Centre Universitaire d'Orsay, 91405 Orsay Cedex,2 France

Received 10 March 2000/Accepted 12 May 2000

Malic enzyme is one of at least five enzymes, known to be present in Corynebacterium glutamicum, capable of carboxylation and decarboxylation reactions coupling glycolysis and the tricarboxylic acid cycle. To date, no information is available concerning the physiological role of the malic enzyme in this bacterium. The malE gene from C. glutamicum has been cloned and sequenced. The protein encoded by this gene has been purified to homogeneity, and the biochemical properties have been established. Biochemical characteristics indicate a decarboxylation role linked to NADPH generation. Strains of C. glutamicum in which the malE gene had been disrupted or overexpressed showed no detectable phenotype during growth on either acetate or glucose, but showed a significant modification of growth behavior during lactate metabolism. The wild type showed a characteristic brief period of exponential growth on lactate followed by a linear growth period. This growth pattern was further accentuated in a malE-disrupted strain (Delta malE). However, the strain overexpressing malE maintained exponential growth until all lactate had been consumed. This strain accumulated significantly larger amounts of pyruvate in the medium than the other strains.

* Corresponding author. Mailing address: Laboratoire de Biotechnologie Bioprocedes, Centre de Bioingénierie Gilbert Durand, Institut National des Sciences Appliquées, 135 Ave. de Rangueil, 31077 Toulouse Cedex 4, France. Phone: (33) 561 559 489. Fax: (33) 561 559 400. E-mail: lindley{at}insa-tlse.fr.

Applied and Environmental Microbiology, July 2000, p. 2981-2987, Vol. 66, No. 7
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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