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.
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.
Murakami
S
S
Department of Bio-functional Chemistry, Kobe University, Nada-ku, Japan.
Kodama
N
N
Shinke
R
R
Aoki
K
K
D000595:Amino Acid Sequence; D001173:Arthrobacter / Q000235:genetics; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D050548:Catechol 1,2-Dioxygenase; D002396:Catechols / Q000378:metabolism; D003001:Cloning, Molecular; D049308:Dioxygenases; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D015723:Gene Library; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010105:Oxygenases / Q000145:classification* / Q000235:genetics* / Q000378:metabolism; D015183:Restriction Mapping; D012240:Rhodococcus / Q000201:enzymology* / Q000235:genetics*; D012270:Ribosomes / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity / Q000235:genetics; D014170:Transformation, Genetic
D002396:Catechols; C031136:3-methylcatechol; C018599:4-methylcatechol; D010105:Oxygenases; D049308:Dioxygenases; D050548:Catechol 1,2-Dioxygenase