A PURIFICATION METHOD FOR SPECIFIC SERINE PROTEASES USING ONE-STEP AFFINITY-CHROMATOGRAPHY

A simple affinity chromatography procedure for specific isolation of s erine proteases is described. The procedure was tested using enzymes f rom five microbial and one plant source. Feather keratin, covalently b ound to controlled-pore glass, was the support and magnesium chloride was used in the elution buffers instead of zinc chloride. This enabled one-step isolation of serine proteases present in the biological mate rials used. The small (15 cm x 1 cm) controlled-pore keratin-glass col umn allowed high flow rates and protected the proteases from autodiges tion during the chromatography process. The serine proteases were elut ed from the column with good recovery (40-84-6%) and a purification ef ficiency between 5 and 7. The purified proteases were homogeneous by p olyacrylamide gel electrophoresis.