Methyl-coenzyme M reductase (MCR) is the key enzyme in the methane formation step of the methanogenesis pathway. MCR catalyzes the formation of methane from methyl-coenzyme M (CH3-S-CoM) and coenzyme B (HS-CoB) in methanogenic archaea. It has been shown that MCR is also able to drive the methane oxidation reaction. We showed functional expression of Methyl-coenzyme M reductase (MCR) from methanothermobacter marburgensis in methanosarcina acetivorans in both transcriptional and translational level. The methane formation yield (mol methane/mol biomass) has been increased by 60% in the recombinant strain expressing M. marburgensis MCR compared to the wild-type M. acetivorans strain. Nanoluciferase has been expressed in M. acetivorans as a reporter system with up to 1000 times more activity over the background. MCR is a heterohexamer that has two sets of α,β,γ subunits with two active site nickel-containing F430 cofactors. Random mutagenesis of α subunit (mcrA) of M. marburgensis MCR has been used to engineer MCR activity in methanogenesis pathway.

Engineering anaerobic methanogenic pathway and methyl-coenzyme M reductase in Methanosarcina acetivorans

251st National Meeting (2016)

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