Characterization of a Corynebacterium glutamicum lactate utilization operon induced during temperature-triggered glutamate production

Stansen, K.; Delaunay, S.; Wendisch, V. F.; Eggeling, L.; Uy, D.; Goergen, J. L.

doi:10.1128/AEM.71.10.5920-5928.2005

Journal Article

Characterization of a Corynebacterium glutamicum lactate utilization operon induced during temperature-triggered glutamate production

Stansen, K.FZJ* ; Uy, D. ; Delaunay, S. ; Eggeling, L.FZJ* ; Goergen, J. L. ; Wendisch, V. F.FZJ*

2005
Soc. Washington, DC [u.a.]

Applied and environmental microbiology 71, 5920 - 5928 (2005) [10.1128/AEM.71.10.5920-5928.2005]

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Please use a persistent id in citations: http://hdl.handle.net/2128/2422 doi:10.1128/AEM.71.10.5920-5928.2005

Abstract: Gene expression changes of glutamate-producing Corynebacterium glutamicum were identified in transcriptome comparisons by DNA microarray analysis. During glutamate production induced by a temperature shift, C. glutamicum strain 2262 showed significantly higher mRNA levels of the NCgl2816 and NCgl2817 genes than its non-glutamate-producing derivative 2262NP. Reverse transcription-PCR analysis showed that the two genes together constitute an operon. NCgl2816 putatively codes for a lactate permease, while NCgl2817 was demonstrated to encode quinone-dependent l-lactate dehydrogenase, which was named LldD. C. glutamicum LldD displayed Michaelis-Menten kinetics for the substrate l-lactate with a K(m) of about 0.51 mM. The specific activity of LldD was about 10-fold higher during growth on l-lactate or on an l-lactate-glucose mixture than during growth on glucose, d-lactate, or pyruvate, while the specific activity of quinone-dependent d-lactate dehydrogenase differed little with the carbon source. RNA levels of NCgl2816 and lldD were about 18-fold higher during growth on l-lactate than on pyruvate. Disruption of the NCgl2816-lldD operon resulted in loss of the ability to utilize l-lactate as the sole carbon source. Expression of lldD restored l-lactate utilization, indicating that the function of the permease gene NCgl2816 is dispensable, while LldD is essential, for growth of C. glutamicum on l-lactate.

Keyword(s): Bacterial Proteins: genetics (MeSH) ; Bacterial Proteins: metabolism (MeSH) ; Corynebacterium glutamicum: genetics (MeSH) ; Corynebacterium glutamicum: metabolism (MeSH) ; Gene Expression Regulation, Bacterial (MeSH) ; Glutamic Acid: metabolism (MeSH) ; Lactic Acid: metabolism (MeSH) ; Mutation (MeSH) ; Oligonucleotide Array Sequence Analysis (MeSH) ; Operon (MeSH) ; Proteome (MeSH) ; Temperature (MeSH) ; Transcription, Genetic (MeSH) ; Bacterial Proteins ; Proteome ; Lactic Acid ; Glutamic Acid ; J