Improvement in the behavior of bromelain coupled to pNIPAm polymers containing acrylamide or acrylic acid

Bromelain was coupled to N-isopropyl acrylamide (NIPAm)polymers, synthesized using NIPAm and various concentrations of acrylamide (Ac) or acrylic acid (AAc). Incorporation of Ac/AAcinto the polymer increased the LCST (lower critical solutiontemperature) in a concentration dependent manner but AAc wasmore effective in this regard; the LCST rose to 40°C when 6 percent AAc was used. Incorporation of Ac/AAc increased the coupling of enzyme to the polymer and the η (effectiveness factor) of the coupled enzyme, moderately. Various studies indicate that such incorporation of hydrophilic monomers into the polymer does not impair its capacity to couple enzyme or expression of the activity of bound enzyme but seems to actually improve the stability of the enzyme against heat induced inactivation and alkaline pH.