Direct Hydroxylation of Benzene to Phenol by Cytochrome P450BM3 Triggered by Amino Acid Derivatives

Abstract

The selective hydroxylation of benzene to phenol, without the formation of side products resulting from overoxidation, is catalyzed by cytochrome P450BM3 with the assistance of amino acid derivatives as decoy molecules. The catalytic turnover rate and the total turnover number reached 259 min−1 P450BM3−1 and 40 200 P450BM3−1 when N‐heptyl‐l‐proline modified with l‐phenylalanine (C7‐l‐Pro‐l‐Phe) was used as the decoy molecule. This work shows that amino acid derivatives with a totally different structure from fatty acids can be used as decoy molecules for aromatic hydroxylation by wild‐type P450BM3. This method for non‐native substrate hydroxylation by wild‐type P450BM3 has the potential to expand the utility of P450BM3 for biotransformations.

Molecular Design and Regulation of Metalloenzyme Activities through Two Novel Approaches: Ferritin and P450s

“…Based on the consideration ii), we synthesized thirdgeneration fluoride free decoy molecules ( Figure 18). 80 As expected, the benzene hydroxylation proceeded in good yields in the presence of the third-generation decoy molecules up to 260 turnovers (min ¹1 P450BM3 ¹1 ) ( Table 5).…”

Rational and semi-rational engineering of cytochrome P450s for biotechnological applications

“…The crystal structures of P450BM3 bound to different substrates have showed that this residue is positioned closely to the entrance of the substrate access channel. Interestingly, the side chain of Arg47 actually faces away from the active site and has not been observed to form a hydrogen bond with the substrate in the N -palmitoylglycine-bound, N -palmitoylmethionine-bound or C7- L -Pro- L -Phe (a decoy molecule)-bound crystal structures [ [52] , [53] , [54] ]. However, Arg47 is undoubtedly important for substrate binding, as the R47S mutation has been shown to enhance the binding selectivity of this enzyme towards N -acyl homoserine lactone and its derivatives by nearly 250-fold [ 55 , 56 ].…”

Homogeneous Oxygenase Catalysis

“…The coupling efficiency of this transformation reached up to 43% (Scheme 28). 119 Further screening of carboxylic acids and amino acids as efficient decoy molecules is expected to enhance the catalytic activity of P450 BM3 . This work represents an attractive alternative for phenol production and shows that amino acid derivatives with a totally different structure from fatty acids can also be used as decoy molecules for aromatic hydroxylation by WT P450 BM3 .…”