Use of chondroitin sulphate lyases in scar treatment: kinetic considerations of macromolecular crowding

Abstract

Chondroitin sulphate lyases ABC (ChABCs) have been exploited to digest upregulated chondroitin sulphate in nerve injury and disc herniation. The decay of enzyme activity under in vivo conditions remains an issue. In vitro, ChABCs showed activity decline by 60% to 70% at 37 oC in 30 min. By supplementation with Ficoll 400, the ChABC activity was sustainable for at least the 30-min assay period. Time-resolved UV absorption spectroscopy indicated the stabilisation of protein conformation under Ficoll supplementation at 200 mg/ml. The crowdedness led to lowering the Vmax and Km of the enzyme. At chondroitin sulphate C concentration as high as 1 mg/ml, enzyme activity was maximal when Ficoll supplementation was at 50 mg/ml for exolyase or 100 mg/ml for endolyase. Our data offer macromolecular crowding as means to extend the half-life of the lyases.