Single step purification of methylenetetrahydromethanopterin reductase from Methanobacterium thermoautotrophicum by specific binding to Blue Sepharose CL‐6B

摘要

>Methylenetetrahydromethanopterin reductase from metanogenic archaebacteria catalyzes the reversible reduction of N 5,N 10-methylenetetrahydro-methanopterin to N 5-methyltetrahydromethanopterin with reduced coenzyme F₄₂₀ as electron donor. The enzyme is involved in methane formation from CO₂, and in methanoi disproportionation to CO₂ and CH₄. We report here that the reductase from Methanobacterium thermoautotrophicum specifically binds to Blue Sepharose CL-6B. Binding was competitive with coenzyme F₄₂₀ rather than with NAD, NADP, FAD, FMN, AMP, ADP and ATP. The reductase could also be desorbed with salt. Based on this property an affinity Chromatographie procedure for the purification of the enzyme was developed.