Organic solvent functional group effect on enzyme inactivation by the interfacial mechanism

We have used a bubble column apparatus to study interfacial inactivation of enzymes. The amount of enzyme inactivated was proportional to the area of organic solvent exposed, as is characteristic of the interfacial mechanism. Tests were made with a series of 12 solvents of log P close to 4.0, but with different functional groups. With α- and β-chymotrypsin, inactivation was much less severe with amphiphilic molecules like decyl alcohol, than with less polar compounds (heptane as the extreme case). This corresponds to a correlation with aqueous–organic interfacial tension, and presumably reflects a more polar interface as seen by the enzyme adsorbing from the aqueous phase. A 50% mixture of decyl alcohol and heptane behaved similarly to pure decyl alcohol, which would be expected to accumulate at the interface. With pig liver esterase, the correlation was rather weak, however. Accumulated data for interfacial inactivation by alkanes was examined for the above enzymes, and also papain, trypsin, urease and ribonuclease. The differing sensitivities did not show a clear correlation with any enzyme property, although there was some relationship to adiabatic compressibility, thermal denaturation temperature and mean hydrophobicity.