The role of conformational dynamics in the activity of polymer-conjugated CalB in organic solvents.

Perennial interest in enzyme catalysis has been expanding its applicability from aqueous phases where enzymes are naturally evolved to organic solvents in which the majority of industrial chemical syntheses are carried out. Although conjugating an enzyme with a soluble polymer has been attempted to enhance enzyme activity in organic solvents, the underlying mechanism remains poorly understood in terms of the conformational dynamics and enzyme activity. Herein, we combine LF-NMR measurements and MD simulations to investigate the effects of polymer grafting on the conformational dynamics of CalB in organic solvents and the consequential impacts on the catalytic kinetics, using the lipase-catalyzed transesterification reaction as a model system. LF-NMR measurements confirm that conjugation with a soluble polymer improves the enzyme flexibility in organic solvents, leading to an increase in the catalytic efficiency of up to two orders of magnitude. MD simulations suggest that the conjugated enzyme samples a larger conformational space, compared to its native counterpart, validating the hypothesis that polymer motion enhances enzyme dynamics. These experimental and simulation results provide new insights for enhancing enzyme conformational dynamics and thereby catalytic kinetics in organic solvents.

Graphene oxide enabled long-term enzymatic transesterification in an anhydrous gas flux.

Gas-phase enzymatic catalysis has been long pursued but not yet utilized in industrial processes due to many limitations. Herein, we report a hydroxyl-rich graphene oxide (GO) aerogel that can preserve the enzymatic activity and stability in an anhydrous gas flow by providing a water-like microenvironment. Lipase immobilized in the GO aerogel exhibits a 5 to 10-fold increase in apparent activity than the lyophilized lipase powder in transesterification of geraniol and vinyl acetate in the gas phase and maintains the initial activity for more than 500 h. The solid-state circular dichroism measurement confirms that the lipase keeps its native conformation in the aerogel, and the thermogravimetric analysis shows that water molecules essential for the lipase activity can be replaced by the hydroxyl groups at the GO surface. The versatility of this method is demonstrated for two other lipases with different structures, promising unprecedented applications of enzyme-GO aerogels to gas-phase enzymatic catalysis.