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.
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.
Ras
J
J
Department of Microbiology, Biological Laboratory, Vrije Universiteit, Amsterdam, The Netherlands.
Van Ophem
P W
PW
Reijnders
W N
WN
Van Spanning
R J
RJ
Duine
J A
JA
Stouthamer
A H
AH
Harms
N
N
D000445:Aldehyde Oxidoreductases / Q000235:genetics* / Q000302:isolation & purification; D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D005557:Formaldehyde / Q000378:metabolism*; D005798:Genes, Bacterial / Q000235:genetics*; D000432:Methanol / Q000378:metabolism*; D008969:Molecular Sequence Data; D016296:Mutagenesis; D010231:Paracoccus denitrificans / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D005557:Formaldehyde; C020145:formaldehyde dehydrogenase (glutathione); D000445:Aldehyde Oxidoreductases; D000432:Methanol