AUBURGER, G and WINTER, J (1992) PURIFICATION AND CHARACTERIZATION OF BENZOYL-COA LIGASE FROM A SYNTROPHIC, BENZOATE-DEGRADING, ANAEROBIC MIXED CULTURE. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 37 (6). pp. 789-795. ISSN 0175-7598,
Full text not available from this repository.Abstract
The benzoyl-CoA ligase from an anaerobic syntrophic culture was purified to homogeneity. It had a molecular mass of around 420 kDa and consisted of seven or eight subunits of 58 kDa. The temperature optimum was 37-40-degrees-C, the optimum pH around 8.0 and optimal activity required 50-100 mM TRIS-HCl buffer, pH 8.0 and 3-7 mM MgCl2; MgCl2 in excess of 10 mM was inhibitory. The activation energy for benzoate was 11.3 kcal/mol. Although growth occurred only with benzoate as a carbon source, the benzoyl-coenzyme A (CoA) ligase formed benzoyl-CoA esters with benzoate, 2-, 3- and 4-fluorobenzoate, picolinate, nicotinate and isonicotinate. Acetate was activated to acetyl-CoA by an acetyl-CoA synthetase. The K(m) values for benzoate, 2-, 3- and 4-fluorobenzoate were 0.04, 0.28, 1.48 and 0.32 mM, the V(max) values 1.05, 1.0, 0.7 and 0.98 units (U)/mg, respectively. For reduced CoA (CoA-SH) a K(m) of 0.07 mM and a V(max) of 1.05 U/mg and for ATP a K(m) of 0. 16 mM and a V(max) of 1.08 U/mg was determined. Benzoate activation was inhibited by more than 6 mM ATP, presumably by pyrophosphate generation from ATP. The inhibition constant (K(i)) for pyrophosphate was 5.7 mM. No homology of the N-terminal amino acid sequence with that of a 2-aminobenzoyl-CoA ligase of a denitrifying Pseudomonas sp. was found.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | DENITRIFYING PSEUDOMONAS STRAIN; ISOTOPE EXCHANGE; DEGRADATION; PHENOL; CARBOXYLATION; BACTERIA; ENZYME; ACID; METABOLISM; INVITRO; |
| Depositing User: | Dr. Gernot Deinzer |
| Last Modified: | 19 Oct 2022 08:44 |
| URI: | https://pred.uni-regensburg.de/id/eprint/54394 |
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