MODIFICATION OF ALPHA-CHYMOTRYPSIN USING A WATER-SOLUBLE PHOTO-FENTON REAGENT

Modification of an enzyme, alpha-chymotrypsin, was examined by using a water-soluble photo-Fenton reagent. By photoirradiation of the enzyme with the reagent, which can occupy a binding site of the enzyme, a tryptophan residue in the vicinity of the active site was oxidized to N-formylkynurenine. Concurrently, the catalytic properties of the enzyme were largely changed: the K-m was increased and the k(cat) was decreased. The decrease in k(cat) for a specific amide substrate was the most significant among the esters and amides examined. The water-soluble photo-Fenton reagent would be useful to chemically modify relatively Limited regions in biomolecules.