| The Pseudomonas DCSA convening enzyme investigated in this study is probably a monooxygenase enzyme system that converts 3,6-dichlorosalicylic acid to 3,6-dichloro 2,5-dichydroxysalicylic acid (DHSA). The DCSA converting enzyme was purified by a two step procedure that utilized ammonium sulfate fractionation (AS) and gel filtration chromatography. NADH and Mg++ stimulated the activity of the purified enzyme, as well as the activity in whole cells and cell lysates. An estimated MW of 82,000 for the enzyme was obtained by gel filtration. The KM for DCSA of the G-100 purified DCSA converting enzyme is 4.8 × 10−4 M, similar to that measured in cell lysates (2.6 × 10−4 M). The G-100 purified enzyme has maximal activity at 30°C, which is consistent with that in whole cells and cell lysates. The Partially purified enzyme exhibits maximal activity at pH 7.0, which is also consistent with the result in cell lysates. |
