A Simple Purification Method for Chloroperoxidase and Its Use in Organic Media

Abstract

A simple two-step purification method for chloroperoxidase from Caldariomyces fumago has been developed. After filtration of the mycelium the enzyme was bound to a DEAE Sepharose fast flow column. The enzyme was eluted with a 20–200 mM phosphate buffer, pH=5.8. After gel filtration on a Superose 12 HPLC column pure enzyme was obtained. Instead of gel filtration it was also possible to purify the enzyme by concentration over a membrane filter, 10 K cutoff. Concentration to 8% of the original volume yielded an enzyme preparation with Rz=1.31b in 77% yield. The enzyme was active in t-butyl alcohol/water mixtures up to 70% t-butyl alcohol. The sulfoxidation of thioanisole proceeded readily (conversion > 99%) and with high enantioselectivity (>99%) in t-butyl alochol/water mixtures.