ABSTRACT
Poly(maleic anhydride-alt-hexen-1)(poly(MA-alt-H-1)) has been synthesized by radical polymerization and characterized by DSC, FT-IR, acid number determination, viscometric and NMR methods. Data showed that the co-polymer is an alternating co-polymer whose composition does not depend on the monomer feed composition. Invertase was immobilized onto a poly(MA-alt-H-1) membrane via glutaraldehyde and bovine serum albumin. The Km value of poly(MA-alt-H-1)-invertase was approximately 4.4-fold higher than the free enzyme, indicating decreased affinity by the invertase for its substrate (sucrose), whereas Vmax was lower for the immobilized invertase. Immobilization improved the pH stability of the enzyme, as well as its temperature stability. Immobilized samples obtained were stable and could be used many times over a period of 2 months without considerable activity loss.
Subject(s)
Enzymes, Immobilized/chemical synthesis , Enzymes, Immobilized/physiology , Maleic Anhydrides/chemical synthesis , beta-Fructofuranosidase/chemistry , Cross-Linking Reagents/pharmacology , Enzyme Stability , Equipment Reuse , Glutaral/pharmacology , Hydrogen-Ion Concentration , Kinetics , Models, Biological , Osmolar Concentration , Polymers/chemical synthesis , Serum Albumin, Bovine/pharmacology , TemperatureABSTRACT
Amphiphilic bioengineering copolymers having a combination of hydrophilic/hydrophobic linkages and polyelectrolyte behavior, along with an ability to interact with biomacromolecules, in particular with the invertase enzyme, have been synthesized by (a) complex-radical copolymerization of maleic anhydride (MA, the acceptor) and hexene-1 (H-1, the donor) monomers with benzoyl peroxide as the initiator in 1,4-dioxane at 65 degrees C under high-conversion conditions and (b) subsequent grafting (polyesterification) of synthesized poly(MA-alt-H-1) with alpha-methoxy-omega-hydroxy-poly(ethylene oxide) (PEO). Copolymerizations were also carried out in the steady state, in order to essentially reduce the effect of copolymer composition drift. The values of the monomer reactivity ratios (r(1) and r(2)) determined by using the known terminal models of Fineman-Ross (FR) and Kelen-Tüdös (KT), as well as by nonlinear regression (NLR) analysis, are: r(1) = 0.16 and r(2) = 0.30 (FR), r(1) = 0.14 and r(2) = 0.27 (KT), and r(1) = 0.15 and r(2) = 0.29 (NLR), respectively. All the copolymers and graft copolymers were characterized by FTIR spectroscopy, (1)H{(13)C} NMR spectroscopy, viscometric measurements, and chemical (acid number), thermal (DSC and TGA), and X-ray diffraction analyses. Unlike poly(MA-alt-H-1)s, PEO macrobranched graft copolymers exhibit expressed polyelectrolyte and swelling behavior in diluted and concentrated dioxane solutions, respectively. The copolymer and its PEO hyperbranched derivatives can be used as carriers for enzyme immobilization.
