Liang Jiayuan, Zhang Yun, Sun Aijun, Deng Dun, Hu Yunfeng
Key Laboratory of Tropical Marine Bio-resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou, 510301, People's Republic of China.
Guangdong Key Laboratory of Marine Materia Medica, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou, 510301, People's Republic of China.
Appl Biochem Biotechnol. 2016 Feb;178(3):558-75. doi: 10.1007/s12010-015-1894-6. Epub 2015 Oct 14.
A novel microbial esterase BSE01281 identified from the Indian Ocean was cloned, expressed, and functionally characterized. Esterase BSE01281 could enanoselectively resolve (±)-1-phenylethanol and (±)-1-phenylethyl acetate through two types of enzymatic reactions. After the optimization of enzymatic reactions, BSE01281 could efficiently generate (R)-1-phenylethyl acetate with high enantiomeric excess (>99%) and high conversion (42%) after 96 h trans-esterification reactions. Additionally, BSE01281 could also produce (R)-1-phenylethanol (e.e. > 99%) and (S)-1-phenylethyl acetate (e.e. > 95%) at a conversion of 49% through direct hydrolysis of inexpensive racemic 1-phenylethyl acetate for 8 h. Optically pure (R)-1-phenylethanol generated from direct enzymatic hydrolysis of racemic 1-phenylethyl acetate by BSE01281 is not easily prepared by dehydrogenases, which generally follow the "Prelog's rule" and give (S)-1-phenylethanol instead.
从印度洋分离出的新型微生物酯酶BSE01281被克隆、表达并进行了功能表征。酯酶BSE01281可通过两种酶促反应对映选择性拆分（±）-1-苯乙醇和（±）-1-苯乙酸乙酯。经过酶促反应优化后，在96小时的酯交换反应后，BSE01281能够高效生成对映体过量值高（>99%）且转化率高（42%）的（R）-1-苯乙酸乙酯。此外，通过对廉价的外消旋1-苯乙酸乙酯进行8小时的直接水解，BSE01281还能以49%的转化率生成对映体过量值>99%的（R）-1-苯乙醇和对映体过量值>95%的（S）-1-苯乙酸乙酯。通过BSE01281直接酶促水解外消旋1-苯乙酸乙酯生成的光学纯（R）-1-苯乙醇不易通过脱氢酶制备，脱氢酶通常遵循“普雷洛格规则”，生成的是（S）-1-苯乙醇。
