Highly Efficient “Tight Fit” Immobilization of α‐Chymotrypsin in Mesoporous MCM‐41: A Novel Approach Using Precursor Immobilization and Activation

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Abstract
The zymogen α‐chymotrypsinogen A is bound to mesoporous silica MCM‐41 with a protein loading of 170 mg/g solid (MCM‐Z) by a simple stirring in aqueous tris‐HCl buffer (pH 7.2). The bound zymogen is then activated with trypsin to obtain α‐chymotrypsin immobilized on MCM‐41 (MCM‐E.I) that displays an effective enzyme activity corresponding to 65 mg protein/g of solid support (3250 BTEE units/g). A direct immobilization of commercially available α‐chymotrypsin (MCM‐E.II) gives lower loading (1250 BTEE units/g). Protein content of the solid support after immobilization is confirmed by thermogravimetric analysis (TGA). The enzyme is tightly bound to the support and can be used over 100 recycles over 1 week in aqueous as well as reverse micellar media. The immobilized enzyme (MCM‐E.I) has been used for resolution of N‐acetyl‐dl‐amino acid esters and racemic trans‐4‐methoxy‐3‐phenylglycidic acid (PGA) methyl ester.