Chen Jingyi, Huang Shan, Seravalli Javier, Gutzman Howard, Swartz Derrick J, Ragsdale Stephen W, Bagley Kimberly A
Department of Chemistry, Buffalo State, State University of New York, Buffalo, New York 14222, USA.
Biochemistry. 2003 Dec 23;42(50):14822-30. doi: 10.1021/bi0349470.
Carbon monoxide dehydrogenase/acetyl-CoA synthase (CODH/ACS) is a bifunctional enzyme that catalyzes the reversible reduction of carbon dioxide into carbon monoxide and the coupled synthesis of acetyl-CoA from the carbon monoxide produced. Exposure of CODH/ACS from Moorella thermoacetica to carbon monoxide gives rise to several infrared bands in the 2100-1900 cm(-1) spectral region that are attributed to the formation of metal-coordinated carbon monoxide species. Infrared bands attributable to M-CO are not detected in the as-isolated enzyme, suggesting that the enzyme does not contain intrinsic metal-coordinated CO ligands. A band detected at 1996 cm(-1) in the CO-flushed enzyme is assigned as arising from CO binding to a metal center in cluster A of the ACS subunit. The frequency of this band is most consistent with it arising from a terminally coordinated Ni(I) carbonyl. Multiple infrared bands at 2078, 2044, 1970, 1959, and 1901 cm(-1) are attributed to CO binding at cluster C of the CODH subunit. All infrared bands attributed to metal carbonyls decay in a time-dependent fashion as CO(2) appears in the solution. These observations are consistent with the enzyme-catalyzed oxidation of carbon monoxide until it is completely depleted from solution during the course of the experiments.
一氧化碳脱氢酶/乙酰辅酶A合酶(CODH/ACS)是一种双功能酶,可催化二氧化碳可逆还原为一氧化碳,并由产生的一氧化碳偶联合成乙酰辅酶A。将嗜热栖热放线菌的CODH/ACS暴露于一氧化碳会在2100 - 1900 cm(-1)光谱区域产生几个红外波段,这些波段归因于金属配位一氧化碳物种的形成。在刚分离的酶中未检测到归因于M - CO的红外波段,这表明该酶不含有内在的金属配位CO配体。在经CO冲洗的酶中于1996 cm(-1)处检测到的一个波段被指定为由CO与ACS亚基的簇A中的金属中心结合产生。该波段的频率与其由末端配位的Ni(I)羰基产生最为一致。在2078、2044、1970、1959和1901 cm(-1)处的多个红外波段归因于CO与CODH亚基的簇C结合。随着溶液中出现CO2,所有归因于金属羰基的红外波段都以时间依赖的方式衰减。这些观察结果与酶催化一氧化碳氧化直至在实验过程中从溶液中完全耗尽一致。