TY - JOUR
T1 - Optimized over-expression of [FeFe] hydrogenases with high specific activity in Clostridium acetobutylicum
AU - Von Abendroth, Gregory
AU - Stripp, Sven
AU - Silakov, Alexey
AU - Croux, Christian
AU - Soucaille, Philippe
AU - Girbal, Laurence
AU - Happe, Thomas
N1 - Funding Information:
This research work was supported by Süd-Chemie AG, Stiftung der Deutschen Wirtschaft and the Marie Curie fellowship. This work was further supported by the European Commission (7th FP, NEST STRP SOLAR-H2 contract 212508) and the Deutsche Forschungsgemeinschaft (SFB 480).
PY - 2008/11
Y1 - 2008/11
N2 - It was previously shown that Clostridium acetobutylicum is capable to over-express various [FeFe] hydrogenases although the protein yield was low. In this study we report on doubling the yield of the clostridial hydrogenase by replacing the native gene hydA1Ca with a recombinant one via homologous recombination. The purified protein HydAlCa shows an unexpected high specific activity (up to 2257 μmol H2min -1 mg-1) for hydrogen evolution. Furthermore, the highly active green algal hydrogenase HydAlCr from Chlamydomonas reinhardtii was heterologously expressed in C. acetobutylicum, and purified with increased yield (1 mg protein per liter of cells) and high activity (625 nmol H 2 min-1mg-1). EPR studies demonstrate intact H-clusters for homologously and heterologously expressed [FeFe] hydrogenases in the CO-inhibited oxidized redox state, and prove the high efficiency of the C. acetobutylicum expression system.
AB - It was previously shown that Clostridium acetobutylicum is capable to over-express various [FeFe] hydrogenases although the protein yield was low. In this study we report on doubling the yield of the clostridial hydrogenase by replacing the native gene hydA1Ca with a recombinant one via homologous recombination. The purified protein HydAlCa shows an unexpected high specific activity (up to 2257 μmol H2min -1 mg-1) for hydrogen evolution. Furthermore, the highly active green algal hydrogenase HydAlCr from Chlamydomonas reinhardtii was heterologously expressed in C. acetobutylicum, and purified with increased yield (1 mg protein per liter of cells) and high activity (625 nmol H 2 min-1mg-1). EPR studies demonstrate intact H-clusters for homologously and heterologously expressed [FeFe] hydrogenases in the CO-inhibited oxidized redox state, and prove the high efficiency of the C. acetobutylicum expression system.
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U2 - 10.1016/j.ijhydene.2008.07.122
DO - 10.1016/j.ijhydene.2008.07.122
M3 - Article
AN - SCOPUS:55049112510
SN - 0360-3199
VL - 33
SP - 6076
EP - 6081
JO - International Journal of Hydrogen Energy
JF - International Journal of Hydrogen Energy
IS - 21
ER -
