Complete activity profile of Clostridium acetobutylicum [FeFe]-hydrogenase and kinetic parameters for endogenous redox partners.
In Clostridium acetobutylicum, [FeFe]-hydrogenase is involved in hydrogen production in vivo by transferring electrons from physiological electron donors, ferredoxin and flavodoxin, to protons. In this report, by modifications of the purification procedure, the specific activity of the enzyme has been improved and its complete catalytic profile in hydrogen evolution, hydrogen uptake, proton/deuterium exchange and para-H2/ortho-H2 conversion has been determined. The major ferredoxin expressed in the solvent-producing C. acetobutylicum cells was purified and identified as encoded by ORF CAC0303. Clostridium acetobutylicum recombinant holoflavodoxin CAC0587 was also purified. The kinetic parameters of C. acetobutylicum [FeFe]-hydrogenase for both physiological partners, ferredoxin CAC0303 and flavodoxin CAC0587, are reported for hydrogen uptake and hydrogen evolution activities.
Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés, UMR CNRS 5504, UMR INRA 792, INSA, Toulouse, France.
Marie
M
Demuez
Laurent
L
Cournac
Olivier
O
Guerrini
Philippe
P
Soucaille
Laurence
L
Girbal
D046969:Clostridium acetobutylicum / Q000201:enzymology*; D004926:Escherichia coli / Q000235:genetics; D005288:Ferredoxins / Q000737:chemistry* / Q000302:isolation & purification; D005418:Flavodoxin / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D006859:Hydrogen / Q000737:chemistry / Q000378:metabolism; D006864:Hydrogenase / Q000737:chemistry* / Q000302:isolation & purification; D007700:Kinetics; D010084:Oxidation-Reduction
D005288:Ferredoxins; D005418:Flavodoxin; D006859:Hydrogen; D006864:Hydrogenase