Preserving enzymatic activity and enhancing biochemical stability of glutathione transferase by soluble additives under free and tethered conditions.
In the present study, we report the effect of four different soluble additives (sucrose, lactitol, superfloc c577, and dextran sulfate) on the stability of glutathione transferase 1 enzyme from Zea mays (ZmGSTF1-1) under free and tethered conditions at 4 and 25 °C. Among all additives, the best stabilizing effects were observed in the case of superfloc c577 and sucrose at both tested temperatures, yet at distinct concentrations at each condition. Those two stabilizing agents were further combined and potential positive synergistic effects were investigated. In addition, we assessed the long-term storage and operational stability of ZmGSTF1-1 under tethered conditions in the presence of additives, which provided the most conducive effects on its stability under free conditions. Our results strongly suggest that the presence of additives may be beneficial to the stability of the enzyme under both free and tethered conditions. Thermodynamic analysis of the free enzyme in the presence of sucrose, which exhibited the best stabilizing effect at both temperatures, shed light on the possible mechanism of action. Given the considerable importance of the development of GST-based biosensors with prolonged stability, the present work may be of general interest to researchers in the field of applied enzymology.
Laboratory of Enzyme Technology, Department of Biotechnology, School of Food, Biotechnology and Development, Agricultural University of Athens, Athens, Greece.
Christos S
CS
Karamitros
Laboratory of Enzyme Technology, Department of Biotechnology, School of Food, Biotechnology and Development, Agricultural University of Athens, Athens, Greece.
Nikolaos E
NE
Labrou
D003911:Dextrans / Q000494:pharmacology; D004795:Enzyme Stability / Q000187:drug effects; D005079:Excipients / Q000494:pharmacology*; D005982:Glutathione Transferase / Q000737:chemistry* / Q000187:drug effects* / Q000378:metabolism; D007700:Kinetics; D010940:Plant Proteins / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D013395:Sucrose / Q000494:pharmacology; D013402:Sugar Alcohols / Q000494:pharmacology; D003313:Zea mays / Q000201:enzymology
D003911:Dextrans; D005079:Excipients; D010940:Plant Proteins; D013402:Sugar Alcohols; D013395:Sucrose; D005982:Glutathione Transferase; C014635:lactitol
biochemical stability; glutathione transferase; storage stability