Enzyme immobilization on thermosensitive hydrogel microspheres

Toshifumi Shiroya, Nobuko Tamura, Mamoru Yasui, Keiji Fujimoto, Haruma Kawaguchi

Research output: Contribution to journalArticle

75 Citations (Scopus)

Abstract

Precipitation polymerization of N-isopropylacrylamide, acrylamide, and methylenebisacrylamide in water at 70°C resulted in thermosensitive hydrogel microspheres. Carboxyl groups on the microspheres were introduced by hydrolysis, and amino groups by the Hofmann reaction of amide units on the microspheres. Trypsin was immobilized on the carboxylated microspheres using carbodiimide. Phase transitions were detected using a hydrophobic fluorescence probe. The temperatures at which a phase transition occurred were increased by immobilizing enzymes. The enzymatic activity of the immobilized enzymes decreased above the transition temperature. This was attributed to (i) a decrease in the diffusion of substrate; and (ii) entrapment of enzyme in the surface layer of the microspheres. In an attempt to overcome the entrapment, enzymes were immobilized via a hydrophilic spacer (α-(carboxymethyl)-ω-(carboxymethoxy)-poly(oxy-1,2-ethanediyl), PEO acid) to the microspheres. These enzyme-carrying hydrogel microspheres were found to show an enzymatic activity independent of temperature, even though these conjugates show a phase transition at the lower critical solution temperature.

Original languageEnglish
Pages (from-to)267-274
Number of pages8
JournalColloids and Surfaces B: Biointerfaces
Volume4
Issue number5
DOIs
Publication statusPublished - 1995 Jun 23

Fingerprint

Enzyme immobilization
Hydrogel
Microspheres
immobilization
Hydrogels
Immobilization
enzymes
Enzymes
Phase Transition
entrapment
Immobilized Enzymes
Phase transitions
trypsin
Temperature
carboxyl group
spacers
Carbodiimides
amides
temperature
hydrolysis

Keywords

  • Enzyme activity
  • Lower critical solution temperature
  • Thermosensitive hydrogel microsphere

ASJC Scopus subject areas

  • Biotechnology
  • Colloid and Surface Chemistry
  • Physical and Theoretical Chemistry
  • Surfaces and Interfaces

Cite this

Enzyme immobilization on thermosensitive hydrogel microspheres. / Shiroya, Toshifumi; Tamura, Nobuko; Yasui, Mamoru; Fujimoto, Keiji; Kawaguchi, Haruma.

In: Colloids and Surfaces B: Biointerfaces, Vol. 4, No. 5, 23.06.1995, p. 267-274.

Research output: Contribution to journalArticle

Shiroya, Toshifumi ; Tamura, Nobuko ; Yasui, Mamoru ; Fujimoto, Keiji ; Kawaguchi, Haruma. / Enzyme immobilization on thermosensitive hydrogel microspheres. In: Colloids and Surfaces B: Biointerfaces. 1995 ; Vol. 4, No. 5. pp. 267-274.
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