Influence of elevated pressure and pressurized fluids on microenvironment and activity of enzymes

Enzymes have great potential in bioprocess engineering due to their green and mild reaction conditions. However, there are challenges to their application, such as enzyme extraction and purification costs, enzyme recovery, and long reaction time. Enzymatic reaction rate enhancement and enzyme immobilization have the potential to overcome some of these challenges. Application of high pressure (e.g., hydrostatic pressure, supercritical carbon dioxide) has been shown to increase the activity of some enzymes, such as lipases and cellulases. Under high pressure, enzymes undergo multiple alterations simultaneously. High pressure reduces the bond lengths of molecules of reaction components and causes a reduction in the activation volume of enzymesubstrate complex. Supercritical CO2 interacts with enzyme molecules, catalyzes structural changes, and removes some water molecules from the enzyme's hydration layer. Interaction of scCO2 with the enzyme also leads to an overall change in secondary structure content. In the extreme, such changes may lead to enzyme denaturation, but enzyme activation and stabilization have also been observed. Immobilization of enzymes onto silica and zeolite-based supports has been shown to further stabilize the enzyme and provide resistance towards perturbation under subjection to high pressure and scCO2.