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

Author:

Ras J1,Van Ophem P W1,Reijnders W N1,Van Spanning R J1,Duine J A1,Stouthamer A H1,Harms N1

Affiliation:

1. Department of Microbiology, Biological Laboratory, Vrije Universiteit, Amsterdam, The Netherlands.

Abstract

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.

Publisher

American Society for Microbiology

Subject

Molecular Biology,Microbiology

Reference34 articles.

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