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In Situ Studies of Protein Conformation in Supercritical Fluids: Trypsin in Carbon Dioxide
Author(s) -
Zagrobelny Joann,
Bright Frank V.
Publication year - 1992
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp00017a008
Subject(s) - trypsin , supercritical fluid , supercritical carbon dioxide , chemistry , denaturation (fissile materials) , carbon dioxide , fluorescence spectroscopy , in situ , fluorescence , monomer , native state , crystallography , biophysics , enzyme , biochemistry , organic chemistry , nuclear chemistry , biology , physics , polymer , quantum mechanics
The conformation of the monomeric enzyme trypsin has been studied in supercritical carbon dioxide. Steady‐state fluorescence spectroscopy is used to follow the conformation of trypsin in situ as a function of CO 2 density. Our results show for the first time that protein denaturation can occur during the fluid compression step and that the native trypsin is only slightly more stable (1.2 kcal/mol) than the unfolded form. These results demonstrate the power of fluorescence spectroscopy as a tool for studying protein conformation and dynamics in supercritical fluids.