Enzymatic evidence for an involvement of pyruvate dehydrogenase in the anaerobic glycerol metabolism of Klebsiella pneumoniae.
Stoichiometric analysis of pathways involved in anaerobic bioconversion of glycerol by Klebsiella pneumoniae revealed that enzyme(s) in addition to pyruvate formate-lyase (PFL) must be involved in pyruvate decarboxylation. In this work, enzymatic evidence is presented that confirmed a simultaneous involvement of pyruvate dehydrogenase complex (PDH) and excluded the presence of pyruvate:ferredoxin oxidoreductase in this anaerobic bioprocess. The in vitro PDH activity of cell extract from continuous culture was found to be strongly affected by the substrate (glycerol) concentration in medium and cell growth rate (dilution rate). It increases with increasing glycerol concentration and correlates well with the specific substrate uptake rate at different dilution rates in a kind of saturation function. At a similar substrate uptake rate, it decreases with cell growth rate. The in vitro activity of PDH is much higher than its in vivo activity calculated from the pathway stoichiometry but comparable to the calculated in vivo activity of PFL.
Menzel
K
K
GBF-Gesellschaft für Biotechnologische Forschung, Biochemical Engineering Division, Braunschweig, Germany.
Zeng
A P
AP
Deckwer
W D
WD
D000693:Anaerobiosis; D018533:Biomass; D019149:Bioreactors; D003013:Clostridium / Q000378:metabolism; D003470:Culture Media; D005990:Glycerol / Q000378:metabolism*; D007658:Ketone Oxidoreductases / Q000378:metabolism; D007700:Kinetics; D007711:Klebsiella pneumoniae / Q000201:enzymology* / Q000254:growth & development / Q000378:metabolism; D011768:Pyruvate Dehydrogenase Complex / Q000378:metabolism*; D019289:Pyruvic Acid / Q000378:metabolism
D003470:Culture Media; D011768:Pyruvate Dehydrogenase Complex; D019289:Pyruvic Acid; D007658:Ketone Oxidoreductases; C039443:pyruvate-flavodoxin oxidoreductase; D005990:Glycerol