The preparation and properties of ficin chemically attached to carboxymethylcellulose

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Abstract
Purified ficin has been coupled to 4 carboxymethylcellusloses (CM-celluloses) by reaction with their acid azide derivatives. Insoluble products containing 1. 8-4. 7 mg of ficin/ 100 mg of product and retaining 8.0-12.0% of the free enzyme''s esterase activity have been obtained. The amount of bound ficin in these preparations is dependent on the degree of carboxymethyl substitution of the CM-cellulose to which the ficin is attached. A shift of the alkaline limb of the pH-activity curve of ficin when chemically attached to CM-cellulose has been shown. Only a small loss has been observed in the enzymic activity of these products when stored at 2[degree] for 4 months. They are more resistant than free enzyme to heat denaturation. Columns of CM-cellulose-ficin have been packed. The degree of hydrolysis of perfused substrate has been measured for different flow rates through the column. The properties of these derivaties have been discussed.