CYP3A4 Activity in the Presence of Organic Cosolvents, Ionic Liquids, or Water‐Immiscible Organic Solvents

  title={CYP3A4 Activity in the Presence of Organic Cosolvents, Ionic Liquids, or Water‐Immiscible Organic Solvents},
  author={Amandine Chefson and Karine Auclair},
P450 enzymes have attracted the attention of chemists for decades because of their impressive ability to catalyze the hydroxylation of inactivated CH bonds. However, their use for synthesis in aqueous systems is limited. We report here a survey of the activity of purified human CYP3A4 in the presence of organic solvents or ionic liquids. We show that CYP3A4 tolerates only small amounts (<15 %) of water‐miscible organic cosolvents or ionic liquids before its activity toward testosterone drops… 
The activity of human CYP2D6 in low water organic solvents
It is reported here that CYP2D6 colyophilized with trehalose and suspended in n‐decane shows higher activity than in aqueous buffer, and provides an alternative strategy to facilitate the use of this enzyme in synthesis.
Enantioselectivity of D-amino acid oxidase in the presence of ionic liquids
Molecular optimization supported that the protein–ionic liquid interactions modulated the structure of the enzyme, especially in [MIM][COO], leading to relaxation in the enantioselectivity of DAAO.
Chiral Polyol Synthesis Catalyzed by a Thermostable Transketolase Immobilized on Layered Double Hydroxides in Ionic liquids
The particular activity of free and immobilized TKgst in [BMIm][Cl] holds promise to extend the applications of TKGst in other ionic liquids and unusual media in biocatalysis.
Organic solvent stability and long‐term storage of myoglobin‐based carbene transfer biocatalysts
The findings demonstrate the robustness of Mb‐based carbene transferases under operationally relevant conditions and should help guide the application of these biocatalysts for synthetic applications.
Ionic liquid effects on the activity of monooxygenase P450 BM-3
P450 monooxygenases oxidize a broad range of substrates including fatty acids, alcohols, aliphatic and aromatic hydrocarbons which are often poorly soluble in water and have Km values in the
Predictable stereoselective and chemoselective hydroxylations and epoxidations with P450 3A4.
This work demonstrates the utility of chemical auxiliaries to mediate the activity of highly promiscuous enzymes and provides a versatile yet controllable system for regio-, chemo-, and stereoselective oxidations at inactivated C-H bonds.
Applications and Mechanisms of Ionic Liquids in Whole-Cell Biotransformation
This work mainly reviews the applications ofILs in whole-cell biotransformations, and the possible mechanisms of ILs in microbial cell biotranformation are proposed and discussed.
Toward advanced ionic liquids. Polar, enzyme-friendly solvents for biocatalysis
This review covers selected examples of enzyme catalyzed reaction in ionic liquids that demonstrate their advantages and unique properties, and point out opportunities for new applications.
Investigation of Classical Organic and Ionic Liquid Cosolvents for Early-Stage Screening in Fragment-Based Inhibitor Design with Unrelated Bacterial and Human Dihydrofolate Reductases.
It is concluded that the organic cosolvents, methanol, dimethylformamide, and dimethylsulfoxide, offer the most balanced conditions for early-stage compound screening as they maintain sufficient biological activity of both DHFRs while allowing for compound dissolution in the millimolar range.


Enzymatic catalysis in nonaqueous solvents.
Catalytic Hydroxylation in Biphasic Systems using CYP102A1 Mutants
Stability of the monooxygenase under the reaction conditions is quite high as revealed by total turnover numbers of up to 12850 in NADPH-dependent cyclohexane hydroxylation and up to 30000 in NADH-dependent myristic acid oxidation.
Rules for optimization of biocatalysis in organic solvents
General rules for the optimization of different biocatalytic systems in various types of media containing organic solvents are derived by combining data from the literature, and the logarithm of the
Properties and Synthetic Applications of Enzymes in Organic Solvents.
  • Carrea, Riva
  • Chemistry, Biology
    Angewandte Chemie
  • 2000
The thermodynamic, kinetic, spectroscopic, and physical approaches that have been adopted to investigate the factors that affect activity, stability, structure, and selectivity of enzymes in organic solvents are examined.
Laboratory evolution of cytochrome P450 BM‐3 monooxygenase for organic cosolvents
By directed evolution, the activity of P450 BM‐3 in the presence of dimethylsulfoxide (DMSO) and tetrahydrofuran (THF) is improved, achieving increases in specific activity up to 10‐fold in 2% (v/v) THF and 6-fold in 25% DMSO.
Room-temperature ionic liquids as replacements for organic solvents in multiphase bioprocess operations.
Organic solvents are widely used in a range of multiphase bioprocess operations including the liquid-liquid extraction of antibiotics and two-phase biotransformation reactions. There are, however,
Replacement of Natural Cofactors by Selected Hydrogen Peroxide Donors or Organic Peroxides Results in Improved Activity for CYP3A4 and CYP2D6
It is shown here that without mutagenesis, the two studied isoforms can use various hydrogen peroxide donors or organic peroxides such as sodium percarbonate, cumene hydroperoxide, and tert-butylhydroperoxide to catalyze reactions in aqueous media and, Interestingly, product formation and initial rates are increased by as much as twofold compared to the use of the natural cofactors.
Enzymatic Catalysis of Formation of Z‐Aspartame in Ionic Liquid − An Alternative to Enzymatic Catalysis in Organic Solvents
With an initial rate of 1.2 ± 0.1 nmol min−1 mg−1, the first report of enzymatic catalysis in an ionic liquid is presented and the enzyme exhibits outstanding stability, which would normally require immobilization.
Enzyme-catalyzed processes in organic solvents.
Lipases can catalyze several other processes in organic media including esterification, aminolysis, acyl exchange, thiotransesterification, and oximolysis; some of these reactions proceed to an appreciable extent only in nonaqueous solvents.
Progress towards the easier use of P450 enzymes.
The attempts to overcome limitations of biocatalysts using approaches such as mutagenesis, chemical modifications, conditions engineering and immobilization are covered.