Carboxylation reaction catalyzed by 2-oxoglutarate:ferredoxin oxidoreductases from Hydrogenobacter thermophilus.
Hydrogenobacter thermophilus TK-6 is a thermophilic, chemolithoautotrophic, hydrogen-oxidizing bacterium that fixes carbon dioxide via the reductive tricarboxylic acid (rTCA) cycle. 2-Oxoglutarate:ferredoxin oxidoreductase (OGOR) is the key enzyme in this cycle that fixes carbon dioxide. The genome of strain TK-6 encodes at least two distinct OGOR enzymes, termed For and Kor. We report here a method for measuring the carboxylation of succinyl-CoA catalyzed by OGORs. The method involves the in vitro coupling of OGOR with ferredoxin and pyruvate:ferredoxin oxidoreductase from strain TK-6, and glutamate dehydrogenase from Sulfolobus tokodaii. Using this method, we determined both the apparent maximum velocities and the K (m) values of For and Kor for the carboxylation of succinyl-CoA. This is the first reported kinetic analysis of carbon fixation catalyzed by OGOR enzymes from the rTCA cycle.
Yamamoto
Masahiro
M
Department of Biotechnology, University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-8657, Japan.
Ikeda
Takeshi
T
Arai
Hiroyuki
H
Ishii
Masaharu
M
Igarashi
Yasuo
Y
D000214:Acyl Coenzyme A / Q000737:chemistry* / Q000378:metabolism; D001419:Bacteria / Q000201:enzymology* / Q000235:genetics; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D002952:Citric Acid Cycle / Q000502:physiology; D016680:Genome, Bacterial / Q000502:physiology; D007658:Ketone Oxidoreductases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D007700:Kinetics
D000214:Acyl Coenzyme A; D001426:Bacterial Proteins; C012046:succinyl-coenzyme A; D007658:Ketone Oxidoreductases; C023096:2-oxoglutarate synthase