Effects of the overexpression of fructose-1,6-bisphosphate aldolase on fermentation pattern and transcription of the genes encoding lactate dehydrogenase and pyruvate formate-lyase in a ruminal bacterium, Streptococcus bovis.
Whether fructose-1,6-bisphosphate (FBP) triggers the transcriptional regulation of the gene expression of lactate dehydrogenase (LDH) and pyruvate formate-lyase (PFL) in Streptococcus bovis was examined by constructing a recombinant strain that overexpresses FBP aldolase (FBA). When the recombinant strain was grown on glucose, intracellular FBP was much lower as compared to the parent strain, whereas dihydroxyacetone phosphate (DHAP) and d-glyceraldehyde-3-phosphate (GAP) were slightly higher. Intracellular ATP and ADP were slightly lower, but the NADH/NAD(+) ratio was not different. When glucose was replaced by lactose, a less readily utilized substrate, there was no great difference in FBP, DHAP, GAP, or adenine nucleotides. Overexpression of FBA decreased the level of LDH-mRNA, and increased the level of PFL-mRNA. Consequently, FBP concentration was positively related to the LDH-mRNA level and inversely related to the PFL-mRNA level. On the contrary, DHAP and GAP concentrations were positively related to the PFL-mRNA level and inversely related to the LDH-mRNA level. The levels of these mRNA were proportional to the amounts of corresponding enzymes in cells. As a result, the ratio of formate to lactate produced was increased by the overexpression of FBA. From these results, it could be presumed that FBP is involved in the transcriptional control of LDH and PFL synthesis in S. bovis.
Asanuma
Narito
N
Department of Life Science, College of Agriculture, Meiji University, Kawasaki 214-8571, Japan. asanuma@isc.meiji.ac.jp
Yoshii
Takahiro
T
Kikuchi
Masahiro
M
Hino
Tsuneo
T
D000123:Acetyltransferases / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D005285:Fermentation; D005634:Fructose-Bisphosphate Aldolase / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D007770:L-Lactate Dehydrogenase / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins; D012417:Rumen / Q000382:microbiology*; D016985:Streptococcus bovis / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D014158:Transcription, Genetic
D011994:Recombinant Proteins; D007770:L-Lactate Dehydrogenase; D000123:Acetyltransferases; C022560:formate C-acetyltransferase; D005634:Fructose-Bisphosphate Aldolase