Wagner Karla, Hummel Anke, Yang Jianing, Horino Satoshi, Kanomata Kyohei, Akai Shuji, Gröger Harald
Chair of Industrial Organic Chemistry and Biotechnology, Faculty of Chemistry, Bielefeld University, Universitätsstraße 25, 33615, Bielefeld, Germany.
Graduate School of Pharmaceutical Sciences, Osaka University, 1-6, Yamadaoka, Suita, Osaka, 565-0871, Japan.
Chembiochem. 2024 Dec 16;25(24):e202400082. doi: 10.1002/cbic.202400082. Epub 2024 Sep 16.
Chiral tertiary alcohols are important organic compounds in science as well as in industry. However, their preparation in enantiomerically pure form is still a challenge due to their complex structure and steric hindrances compared with primary and secondary alcohols, so kinetic resolution could be an attractive approach. Lipase A from Candida antarctica (CAL-A) has been shown to catalyze the enantioselective esterification of various tertiary alcohols with excellent enantioselectivity but low activity. Here we report a mutagenesis study by rational design to improve CAL-A activity against tertiary alcohols. Single mutants of CAL-A were selected, expressed, immobilized and screened for esterification of the tertiary alcohol 1,2,3,4-tetrahydronaphthalen-1-ol. A double mutant V278S+S429G showed a 1.5-fold higher reaction rate than that of the wild type CAL-A, while maintaining excellent enantioselectivity.
手性叔醇在科学和工业领域都是重要的有机化合物。然而，与伯醇和仲醇相比，由于其复杂的结构和空间位阻，以对映体纯的形式制备它们仍然是一个挑战，因此动力学拆分可能是一种有吸引力的方法。南极假丝酵母脂肪酶A（CAL-A）已被证明能以优异的对映选择性催化各种叔醇的对映选择性酯化反应，但活性较低。在此，我们报告一项通过合理设计进行的诱变研究，以提高CAL-A对叔醇的活性。选择CAL-A的单突变体，进行表达、固定化，并筛选其对叔醇1,2,3,4-四氢萘-1-醇的酯化反应。双突变体V278S+S429G的反应速率比野生型CAL-A高1.5倍，同时保持了优异的对映选择性。
