Functional screening of enzymes and bacteria for the dechlorination of hexachlorocyclohexane by a high-throughput colorimetric assay.

摘要: Two distinct microbial dehalogenases are involved in the first steps of degradation hexachlorocyclohexane (HCH) isomers. The enzymes, LinA and LinB, catalyze dehydrochlorination dechlorination reactions HCH respectively, each with isomer specificities. two enzymes hold great promise for use bioremediation residues contaminated soils, although their kinetics specificities currently limiting. Here we report functional screening a library 700 LinB clones generated from soil DNA improved activity by means high throughput colorimetric assay. assay relies upon visual colour change phenol red an aqueous medium, due to pH drop associated reactions. is performed microplate format using intact cells, making it quick simple perform has sensitivity, dynamic range reproducibility. method been validated quantitative gas chromatographic analysis promising clones, revealing some novel variants both superior degrading activities. Some sphingomonad isolates potentially activities were also identified.