Immobilization of amyloglucosidase on alkylamine derivatives of metal-link-activated inorganic supports

A new process to couple amyloglucosidase (AG) to inorganic supports is described. The technique consists in activating the support with a transition metal salt according to the metal-link method and subsequent amination and linkage of the alkylamine derivative using glutaraldehyde. The various parameters susceptible of influencing the properties of the immobilized enzyme (IME) preparation are investigated. The best result are obtained when 100 mg of 1000-Å controlled porous glass (CPG) are treated with 45 mg of TiCl₄ and the activated carrier aminated using a 10-g/L solution of hexamethylenediamine (HMDA) in carbon tetrachloride (10 Ml/100 mg). Preparation obtained according to the process here described show operational stabilities much superior to those of AG immobilized on the same support by the traditional metal-link method or its variations. The mechanism involved in the preparation of the amino derivative of CPG is proposed.