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Purification and immobilization of rabbit liver aldehyde oxidase
Author(s) -
Angelino Steven A. G. F.,
Müller Franz,
van der Plast Henk C.
Publication year - 1985
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260270409
Subject(s) - chemistry , sepharose , immobilized enzyme , aldehyde , aldehyde oxidase , enzyme , chromatography , yield (engineering) , propionaldehyde , organic chemistry , catalysis , xanthine oxidase , materials science , metallurgy
Aldehyde oxidase (E.C. 1.2.3.1) was isolated from rabbit liver and two potential bioaffinity ligands, i.e., 3‐aminocarbonyl‐1‐benzyl‐6‐methylpyridinium bromide and 3‐aminocarbonyl‐1‐benzyl‐4,6‐dimethylpyridinium chloride, were tested for their applicability in a purification procedure for this enzyme. Various supports and different coupling methods were investigated for the immobilization of aldehyde oxidase. Adsorption to n ‐hexyl‐ and n ‐octylamine‐substituted Sepharose 4B and DEAE Sepharose 6B gave the best retention of aldehyde oxidase activity. The storage stability of free enzyme and enzyme immobilized to n ‐octylamine‐substituted Sepharose 4B was studied in several buffers at pH 7.8 and 9.0. This showed that the stability of immobilized enzyme was much less than that of free enzyme. The apparent operational stability of the immobilized enzyme preparation, however, improved substantially compared to soluble enzyme, although the corresponding product yield is still very poor. Coimmobilization of catalase and/or superoxide dismutase provided no significant increase of the apparent operational stability and product yield. A positive effect on both parameters was found for aldehyde oxidase‐ n ‐alkylamine Sepharose 4B preparations by increasing the amount of enzyme adsorbed per unit weight of support, whereas the productivity of these preparations remained about constant.