Cloning and characterization of two catA genes in Acinetobacter lwoffii K24.
Two novel type I catechol 1,2-dioxygenases inducible on aniline media were isolated from Acinetobacter lwoffii K24. Although the two purified enzymes, CD I1 and CD I2, had similar intradiol cleavage activities, they showed different substrate specificities for catechol analogs, physicochemical properties, and amino acid sequences. Two catA genes, catA1 and catA2, encoding by CD I1 and CD I2, respectively, were isolated from the A. lwoffii K24 genomic library by using colony hybridization and PCR. Two DNA fragments containing the catA1 and catA2 genes were located on separate regions of the chromosome. They contained open reading frames encoding 33.4- and 30.4-kDa proteins. The amino acid sequences of the two proteins matched well with previously determined sequences. Interestingly, further analysis of the two DNA fragments revealed the locations of the catB and catC genes as well. Moreover, the DNA fragment containing catA1 had a cluster of genes in the order catB1-catC1-catA1 while the catB2-catA2-catC2 arrangement was found in the catA2 DNA fragment. These results may provide an explanation of the different substrate specificities and physicochemical properties of CD I1 and CD I2.
Kim
S I
SI
Biomolecule Analysis Group, Korea Basic Science Institute, Taejon.
Leem
S H
SH
Choi
J S
JS
Chung
Y H
YH
Kim
S
S
Park
Y M
YM
Park
Y K
YK
Lee
Y N
YN
Ha
K S
KS
D000150:Acinetobacter / Q000201:enzymology / Q000235:genetics*; D000595:Amino Acid Sequence; D001483:Base Sequence; D050548:Catechol 1,2-Dioxygenase; D002396:Catechols / Q000378:metabolism; D002874:Chromosome Mapping; D002876:Chromosomes, Bacterial; D003001:Cloning, Molecular; D049308:Dioxygenases; D005798:Genes, Bacterial; D007526:Isoelectric Point; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010105:Oxygenases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D013379:Substrate Specificity
D002396:Catechols; D010105:Oxygenases; D049308:Dioxygenases; D050548:Catechol 1,2-Dioxygenase