CLONING AND SEQUENCING OF THE CATECHOL 2,3-DIOXYGENASE GENE OF ALCALIGENES SP KF711

The catechol 2,3-dioxygenase is an aromatic ring-fission enzyme cataly zing the conversion of catechol to 2-hydroxymuconic semialdehyde. A ca techol 2,3-dioxygenase gene has been cloned from chromosomal DNA of Al caligenes sp, KF711, and its sequence was determined. The catechol 2,3 -dioxygenase gene was consisted of 927 nucleotides with ATG initiation codon and TGA termination codon, which can encode a polypeptide of mo lecular weight 35 kDa containing 308 amino acid residues. G+C content of the gene was 58 mol%, and a putative ribosome-binding sequence was identified at about 10 nucleotides upstream from the ATG initiation co don. The sequence of catechol 2,3-dioxygenase from Alcaligenes sp. KF7 11 exhibited 81-92% homology at nucleotide level and 84-92% homology a t amino acid level with those of corresponding enzymes encoded in xylE of TOL plasmid, nahH of NAH7 plasmid, and dmpB of Pseudomonas CF600. (C) 1995 Academic Press, Inc.