Abstract
Formylmethanofuran: tetrahydromethanopterin formyltransferase was purified to electrophoretic homogeneity from cells of Methanobacterium thermoautotrophicum. The enzyme is a tetramer of similar or identical subunits (Mr = 41,000). The equilibrium favors transfer of the formyl group to tetrahydromethanopterin (H4MPT) at physiological pH. The product of formyl transfer by the purified enzyme was shown by a number of criteria to be 5-formyl-H4MPT, as opposed to 10-formyl-H4MPT or 5,10-methenyl-H4MPT. Reconstitution of a portion of the methanogenic C1 cycle was effected by combining purified formyltransferase, methenyl-H4MPT cyclohydrolase, formylmethanofuran, and H4MPT to give methenyl-H4MPT. Additional reconstitution experiments established that the formyltransferase is an essential enzyme for the conversion of carbon dioxide to methane. In conjunction with previously published data (Donnelly, M.I., Escalante-Semerena, J.C., Rinehart, K. L., Jr., and Wolfe, R.S. (1985) Arch. Biochem. Biophys. 242, 430-439), these data substantiate the role of 5-formyl-H4MPT as an intermediate of methanogenesis.
Highlights
Formy1methanofuran:tetrahydromethanopterinformyltransferase was purified to electrophoretic homogeneity from cells of Methanobacterium thermoautotrophicum
Reconstitution of a portion of the methanogenic C1 cycle was effected by combining purified formyltransferase, methenyl-H4MPTcyclohydrolase, formylmethanofuran, and H4MPTto give methenylH4MPT
Terin (H4MPT), unique to methanogens and essentifaolr the reduction of carbon dioxide to methane(4-7),has allowed the identification of other C1-coenzyme intermediates
Summary
Formy1methanofuran:tetrahydromethanopterinformyltransferase was purified to electrophoretic homogeneity from cells of Methanobacterium thermoautotrophicum. Additional reconstitution experiments established that the formyltransferase is an essential enzyme for the conversion of carbondioxide to methane. 242, 430-439), these data substantiate the role of 5-formyl-H4MPT as an intermediate of methanogenesis.
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