Enzymology and maturation of reductive dehalogenases

We have isolated a VC-dehalogenating Dehalococcoides and conducted extensive biochemical, genetic and molecular studies in this microorganism (strain VS), as well as on the catabolic vinyl chloride reductase, VcrA.  Vinyl chloride reduction is catalyzed by membrane-associated VcrA and a membrane integrated VcrB subunit, which serves to associate the VcrA subunit to the outer surface of the cytoplasmic membrane. VcrA contains as typical signature two Fe4-S4 clusters as well as a TAT signal peptide sequence (RRDFMK) for directing the export of the assembled enzyme to outer side of the plasma membrane. In addition, VcrA contains a cobalamin as the catalytically active and essential cofactor, where the cobalamin catalyzes the unusual reduction reaction of the organohalide substrate. We are currently developing a system for heterologous expression and in vitro maturation of VcrA and other reductive dehalogenases to initiate structure-function as well as adaptive evolution studies.

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0 September 21, 2010