Production of catechol from benzoate by the wild strain Ralstonia species Ba-0323 and characterization of its catechol 1,2-dioxygenase.
Ralstonia sp. Ba-0323, a wild strain isolated from soil, produced catechol from benzoate and accumulated it outside the cells. The bacterium produced a maximal amount of catechol (1.6 mg/ml) from 3 mg/ml of sodium benzoate in a 20-h growing culture. The conversion rate of benzoate to catechol was 70% on a molar basis. The catechol production by the resting cells increased in the presence of glycerol, and the maximal amount of catechol produced from 3 mg/ml of sodium benzoate reached 1.9 mg/ml at the conversion rate of 83% after 8 h of incubation. Catechol 1,2-dioxygenase, which catalyzed the ring cleavage of catechol, was purified to homogeneity from a cell extract of Ralstonia sp. Ba-0323 growing on benzoate and characterized. The specific activity of the purified enzyme was much lower than those of the dioxygenases from other microorganisms reported. The Km for catechol of the purified enzyme was much higher than those of other dioxygenases. In addition, the NH2-terminal amino acid sequence of the enzyme was less similar to the other catechol 1,2-dioxygenases than they are to each other.
Wang
C L
CL
Division of Science of Biological Resources, The Graduate School of Science and Technology, Kobe University, Rokko, Japan.
Takenaka
S
S
Murakami
S
S
Aoki
K
K
D000595:Amino Acid Sequence; D001565:Benzoates / Q000378:metabolism*; D050548:Catechol 1,2-Dioxygenase; D002396:Catechols / Q000378:metabolism*; D002455:Cell Division; D049308:Dioxygenases; D016946:Gram-Negative Aerobic Rods and Cocci / Q000254:growth & development / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D008969:Molecular Sequence Data; D010105:Oxygenases / Q000302:isolation & purification* / Q000378:metabolism*; D013379:Substrate Specificity
D001565:Benzoates; D002396:Catechols; D010105:Oxygenases; D049308:Dioxygenases; D050548:Catechol 1,2-Dioxygenase; C034221:catechol