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Microbiology 155 (2009), 1360-1375; DOI  10.1099/mic.0.022004-0
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Microbiology 155 (2009), 1360-1375; DOI  10.1099/mic.0.022004-0
© 2009 Society for General Microbiology

Regulation of ldh expression during biotin-limited growth of Corynebacterium glutamicum

Christiane Dietrich1,3, Aimé Nato1,3, Bruno Bost1,3, Pierre Le Maréchal2,3 and Armel Guyonvarch1,3

1 Université Paris-Sud, IGM, UMR 8621, Orsay F-91405, France
2 Université Paris-Sud, IBBMC, UMR 8619, Orsay F 91405, France
3 CNRS, Orsay F-91405, France

Correspondence
Armel Guyonvarch
armel{at}igmors.u-psud.fr

Corynebacterium glutamicum is a biotin-auxotrophic bacterium and some strains efficiently produce glutamic acid under biotin-limiting conditions. In an effort to understand C. glutamicum metabolism under biotin limitation, growth of the type strain ATCC 13032 was investigated in batch cultures and a time-course analysis was performed. A transient excretion of organic acids was observed and we focused our attention on lactate synthesis. Lactate synthesis was due to the ldh-encoded L-lactate dehydrogenase (Ldh). Features of Ldh activity and ldh transcription were analysed. The ldh gene was shown to be regulated at the transcriptional level by SugR, a pleiotropic transcriptional repressor also acting on most phosphotransferase system (PTS) genes. Electrophoretic mobility shift assays (EMSAs) and site-directed mutagenesis allowed the identification of the SugR-binding site. Effector studies using EMSAs and analysis of ldh expression in a ptsF mutant revealed fructose 1-phosphate as a highly efficient negative effector of SugR. Fructose 1,6-bisphosphate also affected SugR binding.

Abbreviations: EMSA, electrophoretic mobility shift assay; β-Gal, β-galactosidase; Ldh, lactate dehydrogenase; PTS, phosphotransferase system






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