Reactivation of thermally inactivated enzymes by free and immobilized chaperonin GroEL/ES.
Thermally inactivated bovine deoxyribonuclease I (DNase I) and yeast enolase were reactivated by GroEL/ES from Escherichia coli. In both cases, GroEL/ ES was found to have the ability to reactivate inactivated enzymes in an ATP-dependent manner. GroEL/ ES can interact with the enzymes that were denatured at high temperature and convert them to the active conformations. To test the applicability of GroEL/ES to the reactivation processes of thermally inactivated enzymes, GroEL/ES was immobilized using formyl-Cellulofine (GroEL/ES-Cellulofine) and its performance was studied. GroEL/ES-Cellulofine retained a sufficiently high ability to reactivate enzymes. Moreover, GroEL/ES-Cellulofine could be used repeatedly, indicating high durability. These results indicate that immobilized chaperonin is effective for reactivation of enzymes that are thermally inactivated in various bioprocesses.
Teshima
T
T
Department of Chemical Science and Engineering, Faculty of Engineering, Kobe University, Japan.
Kondo
A
A
Fukuda
H
H
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001709:Biotechnology / Q000379:methods; D002417:Cattle; D018835:Chaperonin 10 / Q000235:genetics / Q000378:metabolism*; D018834:Chaperonin 60 / Q000235:genetics / Q000378:metabolism*; D003850:Deoxyribonuclease I / Q000378:metabolism*; D004789:Enzyme Activation; D004795:Enzyme Stability; D006358:Hot Temperature; D010751:Phosphopyruvate Hydratase / Q000378:metabolism*; D011489:Protein Denaturation; D011994:Recombinant Proteins / Q000378:metabolism; D015003:Yeasts / Q000201:enzymology
D018835:Chaperonin 10; D018834:Chaperonin 60; D011994:Recombinant Proteins; D000255:Adenosine Triphosphate; D003850:Deoxyribonuclease I; D010751:Phosphopyruvate Hydratase