Characterization of ligV essential for catabolism of vanillin by Sphingomonas paucimobilis SYK-6.
The vanillin dehydrogenase gene (ligV), which conferred the ability to transform vanillin into vanillate on Escherichia coli, was isolated from Sphingomonas paucimobilis SYK-6. The ligV gene consists of a 1,440-bp open reading frame encoding a polypeptide with a molecular mass of 50,301 Da. The deduced amino acid sequence of ligV showed about 50% identity with the known vanillin dehydrogenases of Pseudomonas vanillin degraders. The gene product of ligV (LigV) produced in E. coli preferred NAD+ to NADP+ and exhibited a broad substrate preference, including vanillin, benzaldehyde, protocatechualdehyde, m-anisaldehyde, and p-hydroxybenzaldehyde, but the activity toward syringaldehyde was less than 5% of that toward vanillin. Insertional inactivation of ligV in SYK-6 indicated that ligV is essential for normal growth on vanillin. On the other hand, growth on syringaldehyde was only slightly affected by ligV disruption, indicating the presence of a syringaldehyde dehydrogenase gene or genes in SYK-6.
Masai
Eiji
E
Department of Bioengineering, Nagaoka University of Technology, Japan. emasai@vos.nagaokaut.ac.jp
Yamamoto
Yuko
Y
Inoue
Tomohiko
T
Takamura
Kazuhiro
K
Hara
Hirofumi
H
Kasai
Daisuke
D
Katayama
Yoshihiro
Y
Fukuda
Masao
M
D000595:Amino Acid Sequence; D001483:Base Sequence; D001547:Benzaldehydes / Q000737:chemistry / Q000378:metabolism*; D002396:Catechols / Q000737:chemistry / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005798:Genes, Bacterial / Q000502:physiology*; D008954:Models, Biological; D008969:Molecular Sequence Data; D015394:Molecular Structure; D008970:Molecular Weight; D009154:Mutation; D009243:NAD / Q000378:metabolism; D016366:Open Reading Frames; D015183:Restriction Mapping; D017386:Sequence Homology, Amino Acid; D020578:Sphingomonas / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity
D001547:Benzaldehydes; D002396:Catechols; D009243:NAD; C005581:protocatechualdehyde; C100058:vanillin; C032175:benzaldehyde