Advantages of Supports Activated with Divinyl Sulfone in Enzyme Coimmobilization: Possibility of Multipoint Covalent Immobilization of the Most Stable Enzyme and Immobilization via Ion Exchange of the Least Stable Enzyme

Vinyl sulfone (VS)–agarose beads were used to develop a new strategy coimmobilize enzymes with very different stabilities, enabling the reuse of most stable immobilized enzyme. Two model combi-biocatalysts prepared. First, trypsin and chymotrypsin multipoint covalently coimmobilized on VS–agarose, support was blocked ethylenediamine, then β-galactosidase from Aspergillus oryzae via anion exchange. Both proteases more than lactase, which could be released triple combi-biocatalyst by incubation in 400 mM ammonium sulfate at pH 7.0. Four cycles high temperature, partial inactivation, release reimmobilization combi-protease biocatalysts performed, maintaining 80% activity 90% chymotrypsin. The second system coimmobilization ficin. Ficin inactivated when VS–agarose supports, but it this aspartic acid. Trypsin acid ficin cation As enzyme ionically exchanged ficin, submitted four as described above similar results.