Growth rate-dependent control in Enterococcus faecalis: effects on the transcriptome and proteome, and strong regulation of lactate dehydrogenase.
Enterococcus faecalis V583 was grown in a glucose-limited chemostat at three different growth rates (0.05, 0.15, and 0.4 h⁻¹). The fermentation pattern changed with growth rate, from a mostly homolactic profile at a high growth rate to a fermentation dominated by formate, acetate, and ethanol production at a low growth rate. A number of amino acids were consumed at the lower growth rates but not by fast-growing cells. The change in metabolic profile was caused mainly by decreased flux through lactate dehydrogenase. The transcription of ldh-1, encoding the principal lactate dehydrogenase, showed very strong growth rate dependence and differed by three orders of magnitude between the highest and the lowest growth rates. Despite the increase in ldh-1 transcript, the content of the Ldh-1 protein was the same under all conditions. Using microarrays and quantitative PCR, the levels of 227 gene transcripts were found to be affected by the growth rate, and 56 differentially expressed proteins were found by proteomic analyses. Few genes or proteins showed a growth rate-dependent increase or decrease in expression across the whole range of conditions, and many showed a maximum or minimum at the middle growth rate (i.e., 0.15 h⁻¹). For many gene products, a discrepancy between transcriptomic and proteomic data were seen, indicating posttranscriptional regulation of expression.
Laboratory of Microbial Gene Technology and Food Microbiology, Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Science, Ås, Norway.
Ibrahim
I
Mehmeti
Ellen M
EM
Faergestad
Martijn
M
Bekker
Lars
L
Snipen
Ingolf F
IF
Nes
Helge
H
Holo
D001426:Bacterial Proteins / Q000378:metabolism*; D018929:Cell Culture Techniques; D013293:Enterococcus faecalis / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000235:genetics / Q000378:metabolism*; D055442:Metabolome / Q000502:physiology*; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics; D012329:RNA, Bacterial / Q000032:analysis; D059467:Transcriptome / Q000502:physiology*
D001426:Bacterial Proteins; D007527:Isoenzymes; D020543:Proteome; D012329:RNA, Bacterial; D007770:L-Lactate Dehydrogenase; C409667:lactate dehydrogenase 1