TY - JOUR A1 - Jain, Surbhi A1 - Katsyv, Alexander A1 - Basen, Mirko A1 - Müller, Volker T1 - The monofunctional CO dehydrogenase CooS is essential for growth of Thermoanaerobacter kivui on carbon monoxide T2 - Extremophiles N2 - Thermoanaerobacter kivui is a thermophilic acetogen that can grow on carbon monoxide as sole carbon and energy source. To identify the gene(s) involved in CO oxidation, the genome sequence was analyzed. Two genes potentially encoding CO dehydrogenases were identified. One, cooS, potentially encodes a monofunctional CO dehydrogenase, whereas another, acsA, potentially encodes the CODH component of the CODH/ACS complex. Both genes were cloned, a His-tag encoding sequence was added, and the proteins were produced from a plasmid in T. kivui. His-AcsA copurified by affinity chromatography with AcsB, the acetyl-CoA synthase of the CO dehydrogenase/acetyl CoA synthase complex. His-CooS copurified with CooF1, a small iron-sulfur center containing protein likely involved in electron transport. Both protein complexes had CO:ferredoxin oxidoreductase as well as CO:methyl viologen oxidoreductase activity, but the activity of CooSF1 was 15-times and 231-times lower, respectively. To underline the importance of CooS, the gene was deleted in the CO-adapted strain. Interestingly, the ∆cooS deletion mutant did not grow on CO anymore. These experiments clearly demonstrated that CooS is essential for growth of T. kivui on CO. This is in line with the hypothesis that CooS is the CO-oxidizing enzyme in cells growing on CO. KW - Biofuels KW - Thermophilic acetogenic bacteria KW - Wood–Ljungdahl pathway KW - Synthesis gas Y1 - 2021 UR - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/69579 UR - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-695793 SN - 1433-4909 N1 - Open Access funding enabled and organized by Projekt DEAL. N1 - We are obliged to the BMBF (ThermoSynCon, grant no.: 031B0857B) for the continuous support. VL - 26 IS - 1, art. 4 SP - 1 EP - 12 PB - Springer CY - Tokyo ER -