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Applied and Environmental Microbiology, September 2000, p. 3773-3777, Vol. 66, No. 9
Department of Agriculture, Meiji University,
Higashimita, Tama-ku, Kawasaki 214-8571, Japan
Received 6 January 2000/Accepted 9 May 2000
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
Km 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.
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Effects of pH and Energy Supply on Activity and
Amount of Pyruvate Formate-Lyase in Streptococcus
bovis
*
Corresponding author. Mailing address: Department of
Agriculture, Meiji University, Higashimita, Tama-ku, Kawasaki 214-8571, Japan. Phone: 81-44-934-7825. Fax: 81-44-934-7825. E-mail:
hino{at}isc.meiji.ac.jp.
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