Derivatives of methanopterin, a coenzyme involved in methanogenesis.

Degradational studies of methanopterin, a coenzyme involved in methanogenesis, are reported. The results of these studies are in full accordance with the proposed structure of methanopterin as N-[1'-(2''-amino-4''-hydroxy-7'' -methyl-6''-pteridinyl)ethyl]-4-[2', 3', 4', 5'-tetrahydroxypent-1'-yl(5'-1'' )O-alpha-ribofuranosyl-5''-phosphoric acid] aniline in which the phosphate group is esterified with alpha-hydroxyglutaric acid. Acid hydrolysis of methanopterin cleaved the 5'----1'' glycosidic bond and yielded a 'hydrolytic product' which was identified as N-[1'-(2''-amino-4''-hydroxy-7'' -methyl-6''-pteridinyl)ethyl]-4-[2', 3', 4', 5'-tetrahydroxypent-1'-yl]aniline. Alkaline permanganate oxidation of methanopterin yielded 7-methylpterin-6-carboxylic acid. Catalytic (or enzymatic) hydrogenation of methanopterin gave a mixture of 6-ethyl-7-methyl-7,8-dihydropterin, 6-ethyl-7-methylpterin and a third compound, named methaniline which was identified as 4-[2', 3', 4', 5'-tetrahydroxypent-1'-yl(5'----1'')O-alpha -ribofuranosyl-5''-phosphoric acid]aniline, in which the phosphate group is esterified with alpha-hydroxyglutaric acid. Methanosarcina barkeri contains a closely related coenzyme called sarcinapterin, which was identified as a L-glutamyl derivative of methanopterin, where the glutamate moiety is attached to the alpha-carboxylic acid group of the alpha-hydroxyglutaric acid moiety of methanopterin via an amide linkage.

Structural elements of methanopterin, a novel pterin present in Methanobacterium thermoautotrophicum.

During short-time labeling experiments, cells of Methanobacterium thermoautotrophicum incorporate a substantial part of 14CO2 in a yellow fluorescent compound (called YFC) [Daniels, L. & Zeikus, J. G. (1978) J. Bacteriol. 136, 75-84]. As the compound was present only in small amounts, its more abundant, metabolic precursor was identified, extracted and purified by column chromatography. The chromophore of this compound is 2-amino-4-hydroxypteridine (pterin) as indicated by its ultraviolet-visible-light absorption and fluorescence properties. Decomposition studies revealed the presence of a number of structural elements, viz. glutamic acid, phosphate and a hexosamine. 1H-NMR and 13C-NMR spectra pointed to the presence of additional, as yet unidentified, elements. The compound is a complex, novel pterin derivative, which we have called methanopterin.