The activity of human CYP2D6 in low water organic solvents

Abstract

P450 enzymes are of high interest for synthetic applications due to their ability to catalyze hydroxylation reactions at inactivated C-H bonds. The low solubility of many substrates in buffer, however, is limiting the applications of P450s. Our recent demonstration that the P450 enzymes CYP2D6 and CYP3A4 can function very well in biphasic solvent systems is one step towards overcoming this drawback, but is not practical when substrates or products are unstable in water, or with water-soluble products. An alternative strategy, which also facilitates enzyme recycling, is to directly resuspend lyophilized enzyme into nearly anhydrous organic solvents. Interestingly, we report here that CYP2D6 colyophilized with trehalose and suspended in n-decane shows higher activity than in aqueous buffer. This study demonstrates the unexpected high tolerance of CYP2D6 to some low water organic solvents and provides an alternative strategy to facilitate the use of this enzyme in synthesis.

Cytochromes P450 as useful biocatalysts: addressing the limitations

“…A separate study conducted using P450 2D6 found that the enzyme, when co-lyophilised with trehalose and suspended in n-decane, displayed higher activity in this near anhydrous solution compared to aqueous media. 105 Exploiting such biphasic reaction systems increases the range of organic solvents which can be employed and potentially allows access to a wider variety of substrates.…”

Lyophilization conditions for the storage of monooxygenases

“…Lyophilization also facilitates the use of enzymes in non-aqueous media (Serdakowski and Dordick, 2008). Optimal conditions for the lyophilization of several P450s have been investigated (Bauer et al, 1994;Blanquet et al, 2005;Chefson et al, 2007;Zhao and Auclair, 2009). For BVMOs research has focused on engineering thermostable BVMOs which would display a better operational stability (Opperman and Reetz, 2010;van Beek et al, 2012).…”