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Dramatically stabilized phosphotriesterase—polymers for nerve agent degradation
Author(s) -
LeJeune Keith E.,
Mesiano Anita J.,
Bower Samuel B.,
Grimsley Janet K.,
Wild James R.,
Russell Alan J.
Publication year - 1997
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/(sici)1097-0290(19970420)54:2<105::aid-bit2>3.0.co;2-p
Subject(s) - immobilized enzyme , chemistry , polymer , solvent , nerve agent , enzyme , prepolymer , enzyme assay , thermal stability , dimethyl sulfoxide , combinatorial chemistry , polyurethane , organic chemistry , acetylcholinesterase
Phosphotriesterase (EC 3.1.8.1) was immobilized within a polyurethane foam matrix during polymer synthesis using a prepolymer synthesis strategy. In addition to retaining greater than 50% of the enzyme specific activity, numerous benefits were incurred upon immobilization. Orders of magnitude increases in storage and thermal stability (net stabilization energy = 12.5 kJ/mol) were observed without the need for enzyme premodification. The immobilized enzyme system was protease resistant and seemed to display no adverse effects from immobilization, such as an alteration of enzyme function. The organic solvent, dimethyl sulfoxide, also exhibited a stabilizing effect on phosphotriesterase enzyme systems over a range of intermediate concentrations. We attribute these effects in part to direct interaction between the aprotic solvent and metal containing residues present at the enzyme's active site. Our data demonstrate that just 2.5 kg of immobilized enzyme may be sufficient to degrade 30,000 tons of nerve agent in just 1 year. © 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 54: 105–114, 1997.