An approach for protein to be completely reversible to thermal denaturation even at autoclave temperatures.
Reversibility of protein denaturation is a prerequisite for all applications that depend on reliable enzyme catalysis, particularly, for using steam to sterilize enzyme reactors or enzyme sensor tips, and for developing protein-based devices that perform on-off switching of the protein function such as enzymatic activity, ligand binding and so on. In this study, we have successfully constructed an immobilized protein that retains full enzymatic activity even after thermal treatments as high as 120 degrees C. The key for the complete reversibility was the development of a new reaction that allowed a protein to be covalently attached to a surface through its C-terminus and the protein engineering approach that was used to make the protein compatible with the new attachment chemistry.
Iwakura
M
M
National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba Central 6, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566, Japan. masa-iwakura@aist.go.jp
Nakamura
D
D
Takenawa
T
T
Mitsuishi
Y
Y
D019943:Amino Acid Substitution; D004355:Drug Stability; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000235:genetics; D006358:Hot Temperature; D006801:Humans; D016297:Mutagenesis, Site-Directed; D011487:Protein Conformation; D011489:Protein Denaturation; D015202:Protein Engineering; D020673:Protein Renaturation; D013762:Tetrahydrofolate Dehydrogenase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
D004800:Enzymes, Immobilized; D013762:Tetrahydrofolate Dehydrogenase