Thermostabilized chemical derivatives of horseradish peroxidase.
Horseradish peroxidase finds a variety of uses in analysis, immunology, organic synthesis, and biosensors. Although moderately stable, its applicability to biosensors and other fields would be greatly enhanced if it could be made yet more stable. Appropriate chemical modification can substantially stabilize enzymes. Here we describe the use of bis-imidates and of bis-succinimides to modify free amino groups of commercial horseradish peroxidase under mild conditions of pH and temperature. Imidates yielded a marginal stabilization. Some of the succinimide derivatives, however, are much more thermostable than the native enzyme. Apparent half-lives indicate stabilizations of 6- to 23-fold, depending on the bis-succinimide used. These modifications preserve the carbohydrate side chains for subsequent reaction or immobilization.
Ryan
O
O
School of Biological Sciences, Dublin City University, Republic of Ireland.
Smyth
M R
MR
Fágáin
C O
CO
D015374:Biosensing Techniques; D003432:Cross-Linking Reagents; D004120:Dimethyl Suberimidate; D004795:Enzyme Stability; D006735:Horseradish Peroxidase / Q000737:chemistry* / Q000378:metabolism; D006358:Hot Temperature; D007700:Kinetics; D013388:Succinimides; D013816:Thermodynamics
D003432:Cross-Linking Reagents; D013388:Succinimides; D004120:Dimethyl Suberimidate; D006735:Horseradish Peroxidase