A Water-insoluble Trypsin Derivative and its Use as a Trypsin Column

Abstract

THE conversion of enzymes into water-insoluble products possessing specific catalytic activity is of interest since such ‘water-insoluble enzymes’ may readily be removed from the reaction mixture, and can be used for the preparation of columns with enzymic activity. If stable, they may be employed repeatedly to induce specific chemical changes in relatively large amounts of substrate. Adsorption techniques were applied to combine enzymes with insoluble carriers¹. These procedures led, however, to partial denaturation and resulted in preparations from which the reversibly adsorbed enzyme was removed while in contact with substrate. In order to strengthen the linkage between enzyme and carrier their attachment by chemical covalent bonds seemed preferable. Such links should obviously be carried out by functional groups non-essential for enzymic activity. Furthermore, the attachment of the enzyme to the carrier by flexible polymeric side-chains would ensure free movement of the catalyst molecules in the reaction mixture.