Lactate dehydrogenase has no control on lactate production but has a strong negative control on formate production in Lactococcus lactis.
A series of mutant strains of Lactococcus lactis were constructed with lactate dehydrogenase (LDH) activities ranging from below 1% to 133% of the wild-type activity level. The mutants with 59% to 133% of lactate dehydrogenase activity had growth rates similar to the wild-type and showed a homolactic pattern of fermentation. Only after lactate dehydrogenase activity was reduced ninefold compared to the wild-type was the growth rate significantly affected, and the ldh mutants started to produce mixed-acid products (formate, acetate, and ethanol in addition to lactate). Flux control coefficients were determined and it was found that lactate dehydrogenase exerted virtually no control on the glycolytic flux at the wild-type enzyme level and also not on the flux catalyzed by the enzyme itself, i.e. on the lactate production. As expected, the flux towards the mixed-acid products was strongly enhanced in the strain deleted for lactate dehydrogenase. What is more surprising is that the enzyme had a strong negative control ( CLDHJF1 =-1.3) on the flux to formate at the wild-type level of lactate dehydrogenase. Furthermore, we showed that L. lactis has limited excess of capacity of lactate dehydrogenase, only 70% more than needed to catalyze the lactate flux in the wild-type cells.
Andersen
H W
HW
Section of Molecular Microbiology, BioCentrum, Technical University of Denmark, Lyngby, Denmark.
Pedersen
M B
MB
Hammer
K
K
Jensen
P R
PR
D005285:Fermentation; D005561:Formates / Q000378:metabolism*; D006019:Glycolysis; D007770:L-Lactate Dehydrogenase / Q000378:metabolism*; D019344:Lactic Acid / Q000378:metabolism*; D013294:Lactococcus lactis / Q000201:enzymology / Q000254:growth & development / Q000378:metabolism*
D005561:Formates; C030544:formic acid; D019344:Lactic Acid; D007770:L-Lactate Dehydrogenase