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1991
Journal Article
Titel
Purification and characterization of 2-halocarboxylic acid dehalogenase II from Pseudomonas spec. CBS 3.
Alternative
Reinigung und Charakterisierung der 2-Halogencarbonsäure-Dehalogenase II aus Pseudomonas spec. CBS 3
Abstract
2-Halocarboxylic acid dehalogenase 2 from Pseudomonas spec. CBS 3 (EC 3.8.1.2), which had been cloned in E. coli Hb 101 was purified to elec- trophoretic homogeneity from crude extracts of E. coli Hb 101 clone 1164. Ammonium sulfate fractionation and three subsequent chromatographic purification steps vielded a pure enzyme in a 230-fold enrichment. The relative molecular masses as determined by gelfiltration on Superose 12 and SDS-polyac-rylamide gel electrophoresis were 64000 Da for the holoenzyme and 29000 Da for the subunit. The isoelectric point, determined by isoelectric focusing, was at pH 6.2. Substrate spezificity towards chlorinated and brominated substrates was limited to short chain monosubstituted 2-halocarboxylic acids. Fluorocompounds were not converted. The reaction proceeded best at a pH above 9.5 and at a reaction temperature of 40-45 Grad C.