Methanol metabolism in thermotolerant methylotrophic Bacillus species

Abstract For a number of years we have tried to isolate versatile methylotrophic bacteria employing the ribulose monophosphate (RuMP) cycle of formaldehyde fixation. Recently this has resulted in the development of techniques for the selective enrichment and isolation in pure culture of Bacillus strains able to grow in methanol mineral medium over a temperature range between 35 and 60°C. At the optimum growth temperatures (50–55°C), these isolates display doubling times between 40 and 80 min. The metabolism of the strains studied is strictly respiratory. Methanol assimilation is exclusively via the RuMP cycle variants with the fructose bisphosphate (FBP) aldolase cleavage and transketolase (TK)/transaldolase (TA) rearrangement. Whole cells were unable to oxidize formate, and no activities of NAD-(in)dependent formaldehyde and formate dehydrogenases were detected. Formaldehyde oxidation most likely proceeds via the so-called dissimilatory RuMP cycle. The initial oxidation of methanol is catalyzed by an NAD-dependent methanol dehydrogenase present as an abundant protein in all strains. The enzyme from Bacillus sp. C1 has been purified and characterized.