Leigh J A, Rinehart K L, Wolfe R S
Biochemistry. 1985 Feb 12;24(4):995-9. doi: 10.1021/bi00325a028.
Methanofuran (carbon dioxide reduction factor) became labeled when incubated in cell extracts of Methanobacterium under hydrogen and 14CO2 in the absence of methanopterin. Proton NMR spectroscopy revealed that a formyl group was bound to the primary amine of methanofuran. [14C]Formylmethanofuran was enzymically converted to 14CH4 in the presence of CH3-S-CoM [2-(methylthio)ethanesulfonic acid], hydrogen, and methanopterin, establishing the formyl moiety as an intermediate in methanogenesis. In the absence of methanopterin, a substantial portion of the formyl label was oxidized to 14CO2 rather than reduced to 14CH4, consistent with a model in which the C1 intermediate is first bound to methanofuran and then to methanopterin, during its reduction. When CH3-S-CoM was replaced by HS-CoM (2-mercaptoethanesulfonic acid), most of the formyl label was oxidized to 14CO2, indicating that methyl group reduction by the CH3-S-CoM methylreductase is required for the conversion of formylmethanofuran to methane.
在没有甲蝶呤的情况下,当在氢气和(^{14}CO_2)存在下于甲烷杆菌的细胞提取物中孵育时,呋喃甲醇(二氧化碳还原因子)被标记。质子核磁共振光谱显示,一个甲酰基与呋喃甲醇的伯胺结合。在存在甲基硫代辅酶M(2-(甲硫基)乙烷磺酸)、氢气和甲蝶呤的情况下,[14C]甲酰呋喃甲醇被酶促转化为(^{14}CH_4),确立了甲酰基部分作为甲烷生成过程中的一个中间体。在没有甲蝶呤的情况下,很大一部分甲酰标记被氧化为(^{14}CO_2),而不是还原为(^{14}CH_4),这与在还原过程中C1中间体首先与呋喃甲醇结合然后与甲蝶呤结合的模型一致。当甲基硫代辅酶M被辅酶M(2-巯基乙烷磺酸)取代时,大部分甲酰标记被氧化为(^{14}CO_2),表明甲酰呋喃甲醇转化为甲烷需要甲基硫代辅酶M甲基还原酶进行甲基还原。