Institute of Biochemistry, Heinrich-Heine University Düsseldorf, Universitätsstr. 1, Düsseldorf 40225, Germany.
Chem Commun (Camb). 2021 Jan 14;57(4):520-523. doi: 10.1039/d0cc06729f. Epub 2020 Dec 17.
Saturation mutagenesis at seven first-sphere residues of the cytochrome P450 monooxygenase 154E1 (CYP154E1) from Thermobifida fusca YX was applied to construct a variant with only three substitutions that enabled the effective two-step synthesis of the potential antidepressant (2R,6R)-hydroxynorketamine. A recombinant E. coli whole-cell system was essential for GC/MS based medium-throughput screening and at the same time facilitated the oxidation of the substrate (R)-ketamine at a higher scale for product isolation and subsequent NMR analysis.
采用饱和突变技术对耐热木霉 CYP154E1（Thermobifida fusca YX）的七个第一圈残基进行突变，构建了一个仅含有三个取代的变体，该变体可有效进行潜在抗抑郁药（2R,6R）-羟基去甲氯胺酮的两步合成。基于 GC/MS 的高通量筛选需要重组大肠杆菌全细胞系统，同时有利于在更大规模上氧化（R）-氯胺酮作为底物，以进行产物分离和随后的 NMR 分析。
