Covalent immobilization of trypsin onto poly(2-hydroxyethyl methacrylate)/polystyrene composite microspheres by cyanogen bromide method and its enzymatic activity

Abstract

Covalent immobilization of trypsin onto poly(2-hydroxyethyl methacrylate)/ polystyrene composite microspheres, produced by emulsifier-free seeded emulsion polymerization technique, was carried out using the cyanogen bromide method under various conditions. The highest enzymatic activities of the trypsin immobilized in this study against N-u-benzoyl-L-arginine ethyl ester and N-ce-benzoyl-L-arginine-p-nitroanilide, as low-molecular substrates and casein as a high-molecular substrate, were high, as corresponding to 80 %, 85 % and 50 % of those of free trypsin, respectively.

The Properties of Covalently Immobilized Trypsin on Soap‐Free P(MMA‐EA‐AA) Latex Particles

“…[26,27] A known amount of the free or the immobilized enzyme was added to 100 ml of 10 À3 M BAEE. Then the pH value was adjusted to 8.0 and maintained at that pH using a pH meter (pHS-25, Shanghai Precision Scientific Instruments Co., Ltd. of China) with 0.02 N KOH under stirring for 10 min.…”

Study on Preparation and Morphology of Uniform Artificial Polystyrene–Poly(methyl methacrylate) Composite Microspheres by Employing the SPG (Shirasu Porous Glass) Membrane Emulsification Technique

“…It is known that surfaces with alternating hydrophobic and hydrophilic domains show excellent biocompatibility (24), because the hydrophobic interaction between carriers and biomaterials or substances in vivo can be weakened. Okubo et al used particles with such morphology, obtained by seeded polymerization, for the immobilization of enzyme and found that high activity of enzyme can be maintained (25,26). Furthermore, the amount of immobilization of biomaterials and affinity between particles and biomaterials can be controlled by adjusting the size and amount of PST hydrophobic or PMMA hydrophilic domain; therefore it can be used in affinity separation of biomaterials.…”

Effect of Lauryl Alcohol on Morphology of Uniform Polystyrene–Poly(methyl methacrylate) Composite Microspheres Prepared by Porous Glass Membrane Emulsification Technique

“…It has been known that a surface with hydrophobic and hydrophilic domains in alternate arrangement showed excellent biocompatibility (24), because the hydrophobic interaction between carriers and biomaterials or substances in vivo can be weakened. Okubo et al used particles with such morphology, obtained by seeded polymerization, for the immobilization of enzyme and found that high enzyme activity can be maintained (25,26). Furthermore, the amount of immobilization of biomaterials and affinity between particles and biomaterials can be controlled by adjusting the size and amount of PST hydrophobic or PMMA hydrophilic domains; therefore it can be used in affinity separation of biomaterials.…”