Karasawa Masayuki, Yonemura Kai, Stanfield Joshua Kyle, Suzuki Kazuto, Shoji Osami
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. 2022 Feb 7;61(7):e202111612. doi: 10.1002/anie.202111612. Epub 2021 Dec 13.
We report an OmpF loop deletion mutant, which improves the cellular uptake of external additives into an Escherichia coli whole-cell biocatalyst. Through co-expression of the OmpF mutant with wild-type P450BM3 in the presence of decoy molecules, the yield of the whole-cell biotransformation of benzene could be considerably improved. Notably, with the decoy molecule C7AM-Pip-Phe the yield duodecupled from 5.7 % to 70 %, with 80 % phenol selectivity. The benzylic hydroxylation of alkyl- and cycloalkylbenzenes was also examined, and with the aid of decoy molecules, propylbenzene and tetralin were converted to 1-hydroxylated products with 78 % yield and 94 % (R) ee for propylbenzene and 92 % yield and 94 % (S) ee for tetralin. Our results suggest that both the decoy molecule and substrate traverse the artificial OmpF channel, synergistically boosting whole-cell bioconversions.
我们报道了一种OmpF环缺失突变体,它能提高外部添加剂进入大肠杆菌全细胞生物催化剂的细胞摄取量。在存在诱饵分子的情况下,通过将OmpF突变体与野生型P450BM3共表达,苯的全细胞生物转化产率可得到显著提高。值得注意的是,使用诱饵分子C7AM-Pip-Phe时,产率从5.7%提高到70%,苯酚选择性为80%。还研究了烷基苯和环烷基苯的苄基羟基化反应,借助诱饵分子,丙苯和四氢萘分别转化为1-羟基化产物,丙苯的产率为78%,对映体过量值(ee)为94%(R),四氢萘的产率为92%,ee为94%(S)。我们的结果表明,诱饵分子和底物都能穿过人工OmpF通道,协同促进全细胞生物转化。