Photoimmobilization of organophosphorus hydrolase within a PEG-based hydrogel

Fotios M. Andreopoulos, Michael J. Roberts, Michael D. Bentley, J. Milton Harris, Eric J. Beckman, Alan J. Russell

Research output: Contribution to journalArticle

40 Scopus citations

Abstract

Organophosphorous hydrolase (OPH) was physically and covalently immobilized within photosensitive polyethylene glycol (PEG)-based hydrogels. The hydroxyl ends of branched polyethylene glycol (b-PEG, four arms, MW = 20,000) were modified with cinnamylidene acetate groups to give water- soluble, photosensitive PEG macromers (b-PEG-CA). The b-PEG-CA macromers underwent photocrosslinking reaction and formed gels upon UV irradiation (>300 nm) in the presence of erythrosin B. Native OPH was pegylated with cinnamylidene-terminated PEG chains (MW = 3400) to be covalently linked with the b-PEG-CA macromers during photogelation. The effect of pegylation on the stability of the enzyme was determined. Furthermore, the effect of enzyme concentration, wavelength of irradiation, and photosensitizer on the stability of the entrapped enzyme was also investigated. The pegylated OPH was more stable than the native enzyme, and the OPH-containing gels exhibited superior stability than the soluble enzyme preparations.

Original languageEnglish (US)
Pages (from-to)579-588
Number of pages10
JournalBiotechnology and Bioengineering
Volume65
Issue number5
DOIs
StatePublished - Dec 5 1999
Externally publishedYes

Keywords

  • Enzyme
  • Hydrogels
  • Immobilization
  • Photocrosslinking

ASJC Scopus subject areas

  • Biotechnology
  • Microbiology

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