The flavin-dependent 2-haloacrylate hydratase (2-HAH) catalyzes conversion of 2-chloroacrylate, a major component in manufacturing of acrylic polymers, to a non-toxic pyruvate through a double bond hydration and subsequent chloride elimination. The enzymes from Burkholderia sp. WS and Pseudomonas sp. YL have been cloned and recombinantly expressed and purified from E. coli. The enzymes contain FAD and are active only in the reduced state, although the reaction is redox neutral. Both enzymes are monomers in solution. Full activity is obtained after incubation of the enzymes with 0.5 mM NAD(P)H. Both enzymes have similar activity with 2-haloacrylate (kcat = ~3.0 s-1; KM = ~10 mM). Interestingly, reaction with NAD(P)H is very slow in the absence of light. When the enzyme is exposed to light, the reduction occurs ~100 fold faster. This apparent light activation is unique to 2-HAHs.  This work was supported by NIH grant R01 GM094469.

Insight into the non-canonical flavin-dependent reaction catalyzed by 2-haloacrylate hydratase

245th National Meeting (2013)

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