Abstract
Formaldehyde dehydrogenase and formate dehydrogenase were purified 45- and 16-fold, respectively, from Hansenula polymorpha grown on methanol. Formaldehyde dehydrogenase was strictly dependent on NAD and glutathione for activity. The K mvalues of the enzyme were found to be 0.18 mM for glutathione, 0.21 mM for formaldehyde and 0.15 mM for NAD. The enzyme catalyzed the glutathine-dependent oxidation of formaldehyde to S-formylglutathione. The reaction was shown to be reversible: at pH 8.0 a K mof 1 mM for S-formylglutathione was estimated for the reduction of the thiol ester with NADH. The enzyme did not catalyze the reduction of formate with NADH. The NAD-dependent formate dehydrogenase of H. polymorpha showed a low affinity for formate (K mof 40 mM) but a relatively high affinity for S-formylglutathione (K mof 1.1 mM). The K mvalues of formate dehydrogenase in cell-free extracts of methanol-grown Candida boidinii and Pichia pinus for S-formylglutathione were also an order of magnitude lower than those for formate. It is concluded that S-formylglutathione rather than free formate is an intermediate in the oxidation of methanol by yeasts.
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Van Dijken, J.P., Oostra-Demkes, G.J., Otto, R. et al. S-formylglutathione: The substrate for formate dehydrogenase in methanol-utilizing yeasts. Arch. Microbiol. 111, 77–83 (1976). https://doi.org/10.1007/BF00446552
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DOI: https://doi.org/10.1007/BF00446552