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Applied and Environmental Microbiology, May 2001, p. 2004-2010, Vol. 67, No. 5
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.5.2004-2010.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Clostridium beijerinckii and Clostridium difficile Detoxify Methylglyoxal by a Novel Mechanism Involving Glycerol Dehydrogenase

Hemachandra Liyanage,1 Shelby Kashket,2 Michael Young,3 and Eva R. Kashket1,*

Department of Microbiology, Boston University School of Medicine, Boston, Massachusetts 021181; The Forsyth Institute, Boston, Massachusetts 021152; and Institute of Biological Sciences, University of Wales, Aberystwyth, Ceredigion SY23 3DD, United Kingdom3

Received 30 October 2000/Accepted 9 February 2001

In contrast to gram-negative bacteria, little is known about the mechanisms by which gram-positive bacteria degrade the toxic metabolic intermediate methylglyoxal (MG). Clostridium beijerinckii BR54, a Tn1545 insertion mutant of the NCIMB 8052 strain, formed cultures that contained significantly more (free) MG than wild-type cultures. Moreover, BR54 was more sensitive to growth inhibition by added MG than the wild type, suggesting that it has a reduced ability to degrade MG. The single copy of Tn1545 in this strain lies just downstream from gldA, encoding glycerol dehydrogenase. As a result of antisense RNA production, cell extracts of BR54 possess significantly less glycerol dehydrogenase activity than wild-type cell extracts (H. Liyanage, M. Young, and E. R. Kashket, J. Mol. Microbiol. Biotechnol. 2:87-93, 2000). Inactivation of gldA in both C. beijerinckii and Clostridium difficile gave rise to pinpoint colonies that could not be subcultured, indicating that glycerol dehydrogenase performs an essential function in both organisms. We propose that this role is detoxification of MG. To our knowledge, this is the first report of targeted gene disruption in the C. difficile chromosome.

* Corresponding author. Mailing address: Department of Microbiology, Boston University School of Medicine, 715 Albany Street, Boston, MA 02118-2526. Phone: (617) 638-4291. Fax: (617) 638-4286. E-mail: ekashket{at}bu.edu.

Applied and Environmental Microbiology, May 2001, p. 2004-2010, Vol. 67, No. 5
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.5.2004-2010.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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