Classification of catechol 1,2-dioxygenase family: sequence analysis of a gene for the catechol 1,2-dioxygenase showing high specificity for methylcatechols from Gram + aniline-assimilating Rhodococcus erythropolis AN-13

• Published in: Gene Vol. 185; no. 1; pp. 49 - 54
• Main Authors: Murakami, Shuichiro, Kodama, Noriko, Shinke, Ryu, Aoki, Kenji
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 1997
• Subjects: Amino Acid Sequence; Aromatic compound; Arthrobacter - genetics; Base Sequence; Binding Sites - genetics; Biodegradation; Catechol 1,2-Dioxygenase; Catechols - metabolism; cis,cis-Muconate; Cloning, Molecular; Dioxygenases; Gene Expression Regulation, Bacterial; Gene Expression Regulation, Enzymologic; Gene Library; Molecular Sequence Data; Open Reading Frames; ortho-Cleavage; Oxygenases - classification; Oxygenases - genetics; Oxygenases - metabolism; Restriction Mapping; Rhodococcus - enzymology; Rhodococcus - genetics; Ribosomes - genetics; Sequence Homology, Amino Acid; Substrate Specificity - genetics; Transformation, Genetic; β-Ketoadipate pathway; β-Ketoadipate pathway; Biodegradation; aa, amino acid(s); C-1, Pseudomonas arvilla C-1; ortho-Cleavage; P34O, protocatechuate 3,4-dioxygenase; ORF, open reading frame; bp, base pair(s); C12O, catechol 1,2-dioxygenase(s); Ac, Acinetobacter calcoaceticus; AN-13, Rhodococcus erythropolis AN-13; mt-2, Pseudomonas putida mt-2; cis,cis-Muconate; RBS, ribosome binding site; catA, gene encoding C12O; nt, nucleotide(s); Pp, Pseudomonas putida; kb, kilobase(s); mA3, Arthrobacter sp. mA3; Aromatic compound; EST1001, Pseudomonas sp. EST1001; C12OII, chlorocatechol 1,2-dioxygenase(s); H12O, hydroxyquinol 1,2-dioxygenase(s)
• ISSN: 0378-1119; 1879-0038
• DOI: 10.1016/S0378-1119(96)00629-4

Gram + aniline-assimilating Rhodococcus erythropolis AN-13 (AN-13) produces catechol 1,2-dioxygenase (C12O) showing high enzymatic activities for 3- and 4-methylcatechols [Aoki et al. (1984) Agric. Biol. Chem. 48, 2087–2095]. A 3.0 kb Sau3AI fragment carrying a gene encoding C12O ( catA) was cloned by selection of transformants showing C12O activity from a gene library of AN-13. Furthermore, we specified a 1.6 kb SalI fragment containing catA from the Sau3AI fragment by subcloning. Sequence analysis revealed that the 1.6 kb SalI fragment carried a 855 bp open reading frame (ORF) encoding the entire AN-13 catA, preceded by a potential ribosome binding site (RBS). From comparison of the deduced amino acid (aa) sequence of C12O from AN-13 with other C12O reported previously, it was found that the AN-13 enzyme shares 56.0% aa sequence identity with C12O from Arthrobacter sp. mA3 (mA3) [Eck and Belter (1991) Gene 123, 87–92] compared with less than 36.4% aa sequence identities with others. In conclusion, we classified all C12O including the AN-13 enzyme into three subfamilies on the basis of similarity of aa sequences, numbers of aa residues, and substrate specificity.