Effects of pH and energy supply on activity and amount of pyruvate formate-lyase in Streptococcus bovis.
The enzyme system of pyruvate formate-lyase (PFL) in Streptococcus bovis was investigated by isolating PFL and PFL-activating enzyme (PFL-AE) from S. bovis, flavodoxin from Escherichia coli, and chloroplasts from spinach. In this study, the PFL and PFL-AE in S. bovis were found to be similar to those in E. coli, suggesting that the activating mechanisms are similar. The optimal pH of S. bovis PFL was 7.5, which is in contrast to the optimal pH of S. bovis lactate dehydrogenase, which is 5.5. The apparent K(m) of S. bovis PFL was 2 mM. The intermediates of glycolysis, dihydroxyacetone phosphate (DHAP) and D-glyceraldehyde-3-phosphate (GAP), were shown to inhibit PFL activity. The concentrations of intracellular DHAP and GAP in S. bovis ranged from 1.9 mM to less than 0.1 mM and from 0.6 mM to less than 0.05 mM, respectively, depending on the energy supply. The wide variations in DHAP and GAP levels indicated that PFL activity is allosterically regulated by these triose phosphates in vivo. The amount of PFL protein, as determined by Western blot analysis with polyclonal antibody, changed in parallel with the level of pfl-mRNA, responding to the culture conditions. These observations confirm that PFL synthesis is regulated at the transcriptional level and support the hypothesis that S. bovis shifts the fermentation pathway from acetate, formate, and ethanol production to lactate production when the pH is low and when excess energy is supplied.
Asanuma
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Department of Agriculture, Meiji University, Higashimita, Tama-ku, Kawasaki 214-8571, Japan.
Hino
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D000123:Acetyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002736:Chloroplasts; D003470:Culture Media; D004099:Dihydroxyacetone Phosphate / Q000378:metabolism / Q000494:pharmacology; D004798:Enzymes / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology; D005418:Flavodoxin / Q000378:metabolism; D005561:Formates / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D005986:Glyceraldehyde 3-Phosphate / Q000378:metabolism / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D016985:Streptococcus bovis / Q000201:enzymology* / Q000254:growth & development
D003470:Culture Media; D004798:Enzymes; D005418:Flavodoxin; D005561:Formates; D005986:Glyceraldehyde 3-Phosphate; D004099:Dihydroxyacetone Phosphate; D000123:Acetyltransferases; C040285:pyruvate formate-lyase activating enzyme; C022560:formate C-acetyltransferase