TY - JOUR
T1 - Characterization of a highly thermostable ß-hydroxybutyryl CoA dehydrogenase from Clostridium acetobutylicum ATCC 824
AU - Sommer, Bettina
AU - Garbe, Daniel
AU - Schrepfer, Patrick
AU - Brück, Thomas
N1 - Funding Information:
This work was supported by the German Federal Ministry of Education and Research (BMBF) through grant No. 0315485B and Clariant, a for-profit company pursuing commercialization of bio-based processes.
PY - 2013
Y1 - 2013
N2 - Higher energy content and hydrophobicity make bio-based n-butanol a preferred building block for chemical and biofuels manufacturing. Butanol is obtained by Clostridium sp. based ABE fermentation process. While the ABE process is well understood, the enzyme systems involved have not been elucidated in detail. The important enzyme ß-hydroxybutyryl CoA dehydrogenase from Clostridium acetobutylicum ATCC 824 (Hbd) was purified and characterized. Surprisingly, Hbd shows extremely high temperature (T > 60 C), pH (4-11) and solvent (1-butanol, isobutanol, ethanol) stability. Hbd catalyzes acetoacetyl CoA hydration to ß-hydroxybutyryl CoA up to pH 9.5, where the reaction is reversed. Substrate (acacCoA, ß-hbCoA) and cofactor (NADH, NAD +, NADPH and NADP+) specificities were determined. We identified NAD+ as an uncompetitive inhibitor. Identification of process relevant enzymes such as Hbd is key to optimize butanol production via cellular or cell-free enzymatic systems.
AB - Higher energy content and hydrophobicity make bio-based n-butanol a preferred building block for chemical and biofuels manufacturing. Butanol is obtained by Clostridium sp. based ABE fermentation process. While the ABE process is well understood, the enzyme systems involved have not been elucidated in detail. The important enzyme ß-hydroxybutyryl CoA dehydrogenase from Clostridium acetobutylicum ATCC 824 (Hbd) was purified and characterized. Surprisingly, Hbd shows extremely high temperature (T > 60 C), pH (4-11) and solvent (1-butanol, isobutanol, ethanol) stability. Hbd catalyzes acetoacetyl CoA hydration to ß-hydroxybutyryl CoA up to pH 9.5, where the reaction is reversed. Substrate (acacCoA, ß-hbCoA) and cofactor (NADH, NAD +, NADPH and NADP+) specificities were determined. We identified NAD+ as an uncompetitive inhibitor. Identification of process relevant enzymes such as Hbd is key to optimize butanol production via cellular or cell-free enzymatic systems.
KW - Butanol production
KW - Clostridium acetobutylicum
KW - Solvent stability
KW - Thermo stability
KW - ß-Hydroxybutyryl CoA dehydrogenase
UR - http://www.scopus.com/inward/record.url?scp=84887029550&partnerID=8YFLogxK
U2 - 10.1016/j.molcatb.2013.10.014
DO - 10.1016/j.molcatb.2013.10.014
M3 - Article
AN - SCOPUS:84887029550
SN - 1381-1177
VL - 98
SP - 138
EP - 144
JO - Journal of Molecular Catalysis B: Enzymatic
JF - Journal of Molecular Catalysis B: Enzymatic
ER -