Abstract
Coenzyme F420-dependent methylenetetrahydromethanopterin dehydrogenase from methanogenic Archaea catalyzes the reversible transfer of a hydride ion from C14a of N5,N10-methylenetetrahydromethanopterin to C5 of coenzyme F420. In this study, we report that this hydride transfer proceeds stereospecifically from the Re face at C14a to the Si face at C5. The results were obtained by using chirally 3H-labelled N5,N10-methylenetetrahydromethanopterin generated via Re-face-specific H2-forming N5,N10-methylenetetrahydromethanopterin dehydrogenase and by analyzing reduced coenzyme F420 via Si-face-specific F420-reducing hydrogenase.

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