Blaut M, Müller V, Fiebig K, Gottschalk G
J Bacteriol. 1985 Oct;164(1):95-101. doi: 10.1128/jb.164.1.95-101.1985.
Methanogenesis from methanol by cell suspensions of Methanosarcina barkeri was inhibited by the uncoupler tetrachlorosalicylanilide. This inhibition was reversed by the addition of formaldehyde. 14C labeling experiments revealed that methanol served exclusively as the electron acceptor, whereas formaldehyde was mainly oxidized to CO2 under these conditions. These data support the hypothesis (M. Blaut and G. Gottschalk, Eur. J. Biochem. 141: 217-222, 1984) that the first step in methanol oxidation depends on the proton motive force or a product thereof. Cell extracts of M. barkeri converted methanol and formaldehyde to methane under an H2 atmosphere. Under an N2 atmosphere, however, formaldehyde was disproportionated to CH4 and CO2, whereas methanol was metabolized to a very small extent only, irrespective of the presence of ATP. It was concluded that cell extracts of M. barkeri are not able to oxidize methanol. In further experiments, the sodium dependence of methanogenesis and ATP formation by whole cells was investigated. Methane formation from methanol alone and the corresponding increase in the intracellular ATP content were strictly dependent on Na+. If, in contrast, methanol was utilized together with H2, methane and ATP were synthesized in the absence of Na+. The same is true for the disproportionation of formaldehyde to methane and carbon dioxide. From these experiments, it is concluded that in M. barkeri, Na+ is involved not in the process of ATP synthesis but in the first step of methanol oxidation.
巴氏甲烷八叠球菌细胞悬浮液由甲醇生成甲烷的过程受到解偶联剂四氯水杨酰苯胺的抑制。添加甲醛可逆转这种抑制作用。14C标记实验表明,在这些条件下,甲醇仅作为电子受体,而甲醛主要被氧化为二氧化碳。这些数据支持了以下假设(M. Blaut和G. Gottschalk,《欧洲生物化学杂志》141: 217 - 222, 1984),即甲醇氧化的第一步取决于质子动力或其产物。巴氏甲烷八叠球菌的细胞提取物在氢气氛围下将甲醇和甲醛转化为甲烷。然而,在氮气氛围下,甲醛歧化为甲烷和二氧化碳,而甲醇仅在很小程度上被代谢,无论是否存在ATP。得出的结论是,巴氏甲烷八叠球菌的细胞提取物无法氧化甲醇。在进一步的实验中,研究了全细胞生成甲烷和形成ATP对钠的依赖性。仅由甲醇生成甲烷以及细胞内ATP含量相应增加严格依赖于钠离子。相反,如果甲醇与氢气一起被利用,则在没有钠离子的情况下也能合成甲烷和ATP。甲醛歧化为甲烷和二氧化碳的情况也是如此。从这些实验可以得出结论,在巴氏甲烷八叠球菌中,钠离子不参与ATP合成过程,而是参与甲醇氧化的第一步。