Nicotinoprotein methanol dehydrogenase enzymes in Gram-positive methylotrophic bacteria

A novel type of alcohol dehydrogenase enzyme has been characterized from Gram-positive methylotrophic (Bacillus methanolicus, the actinomycetes Amycolatopsis methanolica and Mycobacterium gastri) and non-methylotrophic bacteria (Rhodococcus strains). Its in vivo role is in oxidation of methanol and other primary alcohols. B. methanolicus displays activity of an NAD-dependent methanol dehydrogenase (MDH), which is strongly stimulated by a specific (activator) protein. A. methanolica and M. gastri use an N,N'- dimethyl-4-nitrosoaniline (NDMA)-dependent MDH (methanol: NDMA oxidoreductase; MNO). MDH (43 kDa subunit) and MNO (49 kDa subunit) possess similar decameric structures with five-fold symmetry, Both proteins contain Zn2+- and Mg2+-ions and tightly (but non-covalently) bound NAD(P)(H) cofactors. These nicotinoproteins share a high degree of sequence similarity; they belong to Family III of NAD(P)-dependent alcohol dehydrogenases (ADH). A. methanolica extracts also possess dye (dichlorophenol indophenol, DCPIP and 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide, MTT)-linked ADH activities. These represent the overall activities of multi-enzyme systems. MNO is part of the MTT-ADH complex. The other two proteins and their cofactors most likely participate in transfer of reducing equivalents from the NADPH cofactor in MNO to the respiratory chain. (C) 2000 Elsevier Science B.V. All rights reserved.