Abstract
Commercially available poly(vinyl alcohol) (PVA) was cross-linked with terephthalaldehyde, derivatized with 1-fluoro-4-nitrobenzene and reduced by means of sodium dithionite. The carrier obtained, containing N-substituted sulphamic acid groups, was applied to immobilize ribonuclease (E.C. 3.1.4.22). The influence of protein denaturing agents on the immobilization reaction and its pH dependence was investigated. By copolymerizing suitable methacrylates, copolymers with different spacer lengths between the reactive groups and the polymer matrix were synthesized. After their reduction they showed the same swellability and the same content of amino groups. These products were transformed into reactive carriers by diazotization and coupled with ribonuclease, trypsin (E.C. 3.4.21.4), α-chymotrypsin (E.C. 3.4.21.1) and urease (E.C. 3.5.1.5). The amounts of bound proteins and the enzymatic activities of the immobilization products were determined and correlated with the spacer lengths of the applied carriers.
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