areABC genes determine the catabolism of aryl esters in Acinetobacter sp. Strain ADP1.
Acinetobacter sp. strain ADP1 is able to grow on a range of esters of aromatic alcohols, converting them to the corresponding aromatic carboxylic acids by the sequential action of three inducible enzymes: an areA-encoded esterase, an areB-encoded benzyl alcohol dehydrogenase, and an areC-encoded benzaldehyde dehydrogenase. The are genes, adjacent to each other on the chromosome and transcribed in the order areCBA, were located 3.5 kbp upstream of benK. benK, encoding a permease implicated in benzoate uptake, is at one end of the ben-cat supraoperonic cluster for benzoate catabolism by the beta-ketoadipate pathway. Two open reading frames which may encode a transcriptional regulator, areR, and a porin, benP, separate benK from areC. Each are gene was individually expressed to high specific activity in Escherichia coli. The relative activities against different substrates of the cloned enzymes were, within experimental error, identical to that of wild-type Acinetobacter sp. strain ADP1 grown on either benzyl acetate, benzyl alcohol, or 4-hydroxybenzyl alcohol as the carbon source. The substrate preferences of all three enzymes were broad, encompassing a range of substituted aromatic compounds and in the case of the AreA esterase, different carboxylic acids. The areA, areB, and areC genes were individually disrupted on the chromosome by insertion of a kanamycin resistance cassette, and the rates at which the resultant strains utilized substrates of the aryl ester catabolic pathway were severely reduced as determined by growth competitions between the mutant and wild-type strains.
School of Biological Sciences, University of Wales Bangor, Bangor, Gwynedd LL57 2UW, Wales, United Kingdom.
R M
RM
Jones
L S
LS
Collier
E L
EL
Neidle
P A
PA
Williams
D000150:Acinetobacter / Q000201:enzymology* / Q000235:genetics*; D000429:Alcohol Oxidoreductases / Q000235:genetics* / Q000378:metabolism*; D000445:Aldehyde Oxidoreductases / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D002876:Chromosomes, Bacterial; D003001:Cloning, Molecular; D017124:Conserved Sequence; D004926:Escherichia coli; D004950:Esterases / Q000235:genetics* / Q000378:metabolism*; D004952:Esters; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D007700:Kinetics; D008969:Molecular Sequence Data; D010957:Plasmids; D015183:Restriction Mapping; D014158:Transcription, Genetic
D004952:Esters; D000429:Alcohol Oxidoreductases; C027016:benzyl alcohol dehydrogenase; D000445:Aldehyde Oxidoreductases; C027015:benzaldehyde dehydrogenase (NAD+); D004950:Esterases