Department of Chemistry, Graduate School of Science, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8602, Japan.
Core Research for Evolutional Science and Technology (Japan) Science and Technology Agency, 5 Sanbancho, Chiyoda-ku, Tokyo, 102-0075, Japan.
Angew Chem Int Ed Engl. 2018 Sep 17;57(38):12264-12269. doi: 10.1002/anie.201804924. Epub 2018 Jul 23.
An Escherichia coli whole-cell biocatalyst for the direct hydroxylation of benzene to phenol has been developed. By adding amino acid derivatives as decoy molecules to the culture medium, wild-type cytochrome P450BM3 (P450BM3) expressed in E.coli can be activated and non-native substrates hydroxylated, without supplementing with NADPH. The yield of phenol reached 59 % when N-heptyl-l-prolyl-l-phenylalanine (C7-Pro-Phe) was employed as the decoy molecule. It was shown that decoy molecules, especially those lacking fluorination, reached the cytosol of E. coli, thus imparting in vivo catalytic activity for the oxyfunctionalisation of non-native substrates to intracellular P450BM3.
已开发出一种用于苯直接羟化为苯酚的大肠杆菌全细胞生物催化剂。通过在培养基中添加氨基酸衍生物作为诱饵分子，可以激活在大肠杆菌中表达的野生型细胞色素 P450BM3（P450BM3）并使非天然底物羟化，而无需补充 NADPH。当使用 N-庚基-l-脯氨酰-l-苯丙氨酸（C7-Pro-Phe）作为诱饵分子时，苯酚的收率达到 59%。结果表明，诱饵分子，特别是那些缺乏氟化的分子，到达了大肠杆菌的细胞质，从而赋予了细胞内 P450BM3 对非天然底物的氧功能化的体内催化活性。
