Photoimmobilization of organophosphorus hydrolase within a PEG-based hydrogel.
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.
Andreopoulos
F M
FM
Department of Chemical and Petroleum Engineering, 1249 Benedum Hall, University of Pittsburgh, Pittsburgh, Pennsylvania 15261, USA.
Roberts
M J
MJ
Bentley
M D
MD
Harris
J M
JM
Beckman
E J
EJ
Russell
A J
AJ
D043303:Aryldialkylphosphatase; D003432:Cross-Linking Reagents; D004795:Enzyme Stability / Q000528:radiation effects; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000528:radiation effects; D004950:Esterases / Q000737:chemistry / Q000528:radiation effects; D020100:Hydrogels / Q000737:chemistry; D010777:Photochemistry; D011092:Polyethylene Glycols / Q000737:chemistry; D014466:Ultraviolet Rays
D003432:Cross-Linking Reagents; D004800:Enzymes, Immobilized; D020100:Hydrogels; D011092:Polyethylene Glycols; D004950:Esterases; D043303:Aryldialkylphosphatase