Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, 214122 Wuxi, Jiangsu, China.
Bioprocess Biosyst Eng. 2010 Sep;33(7):797-804. doi: 10.1007/s00449-009-0401-2. Epub 2009 Dec 23.
The enantioselective reduction of methyl benzoylformate to (R)-methyl mandelate, an important pharmaceutical intermediate and a versatile resolving agent, was investigated in this study. After minimizing the reaction-specific constraints (constraints dependent on the nature of the substrate and product) by preliminary selection of the reaction parameters, an effective whole cell biocatalyst (Saccharomyces cerevisiae AS2.1392) was obtained by simple screening procedures. Under further optimized conditions, a product concentration of 103 mmol L(-1) could be attained within 5 h with a yield of 85.8% and an enantiometric excess of 95.4%, indicating S. cerevisiae AS2.1392 an efficient biocatalyst for the asymmetric synthesis of (R)-methyl mandelate. Furthermore, resin-based in situ product removal (ISPR) technique was applied to alleviate the substrate and product inhibition or toxicity to the whole cells. The integration of newly isolated biocatalyst and proper ISPR technique provides a practical route for the preparation of optically active pharmaceutical intermediates.
本研究考察了将苯甲酰基甲酸甲酯（一种重要的医药中间体和多功能拆分试剂）对映选择性还原为（R）-甲基扁桃酸。通过初步选择反应参数，最小化了反应特异性约束（取决于底物和产物性质的约束），然后通过简单的筛选程序获得了有效的全细胞生物催化剂（酿酒酵母 AS2.1392）。在进一步优化的条件下，在 5 h 内可以达到 103 mmol L(-1)的产物浓度，产率为 85.8%，对映体过量值为 95.4%，表明酿酒酵母 AS2.1392 是（R）-甲基扁桃酸不对称合成的有效生物催化剂。此外，还应用基于树脂的原位产物去除（ISPR）技术来缓解底物和产物对全细胞的抑制或毒性。新分离的生物催化剂和适当的 ISPR 技术的集成为制备光学活性药物中间体提供了一种实用途径。
