Enzyme Leakage and Multipoint Attachment of Agarose‐Bound Enzyme Preparations

Abstract

The solvolytic detachment of leucine aminopeptidase fromSepharose‐enzyme conjugates with multiple and single anchoring bonds has been studied under a variety of conditions by radiochemical and enzymological methods. The release of the single‐point‐fixed conjugate could be described by a leakage function, derived previously, yielding the first‐order rate constant of the cleavage of the enzyme‐matrix bond. The nucleophile hydroxylamine increased the detachment rate considerably. The release of the immobilized enzyme was incomplete in all experiments even after prolonged times. The enzyme leakage from multipoint‐attached conjugates was still high enough to prohibit a long‐term use of such preparations in routine work at room temperature.