Wang Hui-Hui, Wan Nan-Wei, Da Xin-Yu, Mou Xue-Qing, Wang Zhu-Xiang, Chen Yong-Zheng, Liu Zhi-Qiang, Zheng Yu-Guo
Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310014, China; Key Laboratory of Biocatalysis & Chiral Drug Synthesis of Guizhou Province, Generic Drug Research Center of Guizhou Province, Green Pharmaceuticals Engineering Research Center of Guizhou Province, School of Pharmacy, Zunyi Medical University, Zunyi, China.
Key Laboratory of Biocatalysis & Chiral Drug Synthesis of Guizhou Province, Generic Drug Research Center of Guizhou Province, Green Pharmaceuticals Engineering Research Center of Guizhou Province, School of Pharmacy, Zunyi Medical University, Zunyi, China.
Bioorg Chem. 2023 Sep;138:106640. doi: 10.1016/j.bioorg.2023.106640. Epub 2023 Jun 1.
Enantiopure β-nitroalcohols, as an important class of nitro-containing compounds, are essential building blocks in pharmaceutical and organic chemistry, particularly for the synthesis of β-adrenergic blockers. In this study, we present the successful protein engineering of halohydrin dehalogenase HHDHamb for the enantioselective bio-nitration of various phenyl glycidyl ethers to the corresponding chiral β-nitroalcohols, using the inexpensive, commercially available, and safer nitrite as a nitrating agent. The chiral (R)- and (S)-1-nitro-3-phenoxypropan-2-ols were synthesized by the several enantiocomplementary HHDHamb variants through the whole-cell biotransformation, which showed good catalytic efficiency (up to 43% isolated yields) and high optical purity (up to >99% ee). In addition, we also demonstrated that the bio-nitration method was able to tolerate the substrate at a high concentration of 1000 mM (150 g/L). Furthermore, representative synthesis of two optically active enantiomers of the β-adrenergic blocker metoprolol was successfully achieved by utilizing the corresponding chiral β-nitroalcohols as precursors.
对映体纯的β-硝基醇作为一类重要的含硝基化合物，是药物化学和有机化学中必不可少的构建模块，尤其用于合成β-肾上腺素能阻滞剂。在本研究中，我们展示了卤代醇脱卤酶HHDHamb成功的蛋白质工程改造，该酶使用廉价、市售且更安全的亚硝酸盐作为硝化剂，将各种苯基缩水甘油醚对映选择性生物硝化为相应的手性β-硝基醇。通过几种对映互补的HHDHamb变体，通过全细胞生物转化合成了手性（R）-和（S）-1-硝基-3-苯氧基丙-2-醇，其显示出良好的催化效率（分离产率高达43%）和高光学纯度（高达>99% ee）。此外，我们还证明了生物硝化方法能够耐受1000 mM（150 g/L）的高浓度底物。此外，通过使用相应的手性β-硝基醇作为前体，成功实现了β-肾上腺素能阻滞剂美托洛尔的两种光学活性对映体的代表性合成。
