Enzyme leakage and multipoint attachment of agarose-bound enzyme preparations.
The solvolytic detachment of leucine aminopeptidase from Sepharose-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.
D000596:Amino Acids / Q000032:analysis; D004800:Enzymes, Immobilized / Q000378:metabolism; D007700:Kinetics; D007931:Leucyl Aminopeptidase / Q000378:metabolism; D011485:Protein Binding; D012685:Sepharose; D013056:Spectrophotometry, Ultraviolet
D000596:Amino Acids; D004800:Enzymes, Immobilized; D012685:Sepharose; D007931:Leucyl Aminopeptidase