By site-directed mutagenesis, Cys-116 was converted to Ser-116 in p-hydroxybenzoate hydroxylase (EC 1.14.13.2) from Pseudomonas fluorescens. In contrast to wild-type enzyme, the C116S mutant is no longer susceptible to oxidation by hydrogen peroxide and shows no reactivity towards 5,5'-dithiobis(2-nitrobenzoate). Crystals of the C116S mutant are isomorphous with the crystal form of wild-type enzyme. A difference electron density confirms the mutation made.

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Journal Article | Research Support, Non-U.S. Gov't

Authors

Eschrich K, van Berkel WJ, Westphal AH, de Kok A, Mattevi A, Obmolova G, Kalk KH, Hol WG

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