The Preparation and Some Properties of Bromelain Covalently Attached to O‐(Carboxymethyl)‐Cellulose

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Abstract

Purified bromelain has been attached to CM‐cellulose by reaction with its acid azide derivative to yield insoluble preparations which contain 6.5–11.0 mg of bromelain/100 mg of product. The result of an extensive washing procedure strongly suggests that this protein is attached chemically to the CM‐cellulose. The apparent catalytic constant for the catalysis of the hydrolysis of α‐N‐benzoyl‐l‐arginine ethyl ester (BzArgOEt) at pH 7.0, I= 0.2 and 25.0° by CM‐cellulose‐bromelain is 52% of the corresponding value for the bromelain‐catalysed hydrolysis when the enzyme concentration is estimated by protein analysis and 77% of the value for the bromelain‐catalysed hydrolysis when the enzyme concentration is estimated by analysis of the thiol content of CM‐cellulose‐bromelain. The Michaelis constant for the CM‐cellulose‐bromelain‐catalysed hydrolysis of BzArgOEt is about an order of magnitude lower than the corresponding value for the bromelain‐catalysed hydrolysis at low ionic strength and approaches the value for the bromelain‐catalysed hydrolysis at high ionic strength. The attachment of bromelain to CM‐cellulose protects the enzyme from oxidation at 4° but offers little or no protection from heat denaturation. Copyright © 1968, Wiley Blackwell. All rights reserved

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CITATION STYLE

APA

Wharton, C. W., Crook, E. M., & Brocklehurst, K. (1968). The Preparation and Some Properties of Bromelain Covalently Attached to O‐(Carboxymethyl)‐Cellulose. European Journal of Biochemistry, 6(4), 565–571. https://doi.org/10.1111/j.1432-1033.1968.tb00482.x

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