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J. Bacteriol., Jan 1995, 247-251, Vol 177, No. 1
Copyright © 1995, American Society for Microbiology

Isolation, sequencing, and mutagenesis of the gene encoding NAD- and glutathione-dependent formaldehyde dehydrogenase (GD-FALDH) from Paracoccus denitrificans, in which GD-FALDH is essential for methylotrophic growth

J Ras, PW Van Ophem, WN Reijnders, RJ Van Spanning, JA Duine, AH Stouthamer and N Harms
Department of Microbiology, Biological Laboratory, Vrije Universiteit, Amsterdam, The Netherlands.

NAD- and glutathione-dependent formaldehyde dehydrogenase (GD-FALDH) of Paracoccus denitrificans has been purified as a tetramer with a relative molecular mass of 150 kDa. The gene encoding GD-FALDH (flhA) has been isolated, sequenced, and mutated by insertion of a kanamycin resistance gene. The mutant strain is not able to grow on methanol, methylamine, or choline, while heterotrophic growth is not influenced by the mutation. This finding indicates that GD-FALDH of P. denitrificans is essential for the oxidation of formaldehyde produced during methylotrophic growth.

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