酶-化学级联催化制备(S)-烟碱
Production of(S)-nicotine by enzymatic-chemical cascade catalysis
刘茜琳 1高静 1马丽 1宋浩雷 2贺莹 1郑晓冰1
作者信息
- 1. 河北工业大学 化工学院,天津 300400
- 2. 河北亚诺生物科技有限公司,河北 石家庄 052160
- 折叠
摘要
传统(S)-烟碱合成方法步骤复杂且依赖多种化学催化剂.为了实现(S)-烟碱的绿色合成,设计了酶-化学级联途径:首先,构建了亚胺还原酶(IRED)与甲酸脱氢酶(FDH)的偶联体系,以麦斯明为底物合成(S)-降烟碱;然后,将(S)-降烟碱经Eschweiler-Clarke反应制备(S)-烟碱.通过大肠杆菌宿主分别对IRED和FDH进行表达,得到含酶细胞及酶液,考察了pH和温度对细胞中IRED和FDH酶活力的影响.优化酶催化制备(S)-降烟碱的反应条件,在30℃、pH 7.5条件下,添加质量浓度为30 g/L的麦斯明、600 U/L FDH细胞、900 U/L IRED细胞、0.6 mmol/L NADP+和质量浓度为90 g/L的甲酸钠,(S)-降烟碱的产率>98%,对映体过量(e.e.)值>99%.最后,通过化学法将(S)-降烟碱甲基化得到(S)-烟碱,此步骤产率和e.e.值均达99%以上.
Abstract
The traditional synthesis method of(S)-nicotine involves complex steps and relies on multiple chemical catalysts.Herein,a enzymatic-chemical cascade pathway was designed to achieve the green synthesis of(S)-nicotine.Firstly,a coupling system of imine reductase(IRED)and formate dehydrogenase(FDH)was constructed to catalyzed the synthesis of(S)-nornicotine from myosmine as substrate,then(S)-nicotine was prepared via Eschweiler-Clarke reaction of(S)-nornicotine.The IRED and FDH enzyme-containing cells and enzyme solution were obtained from E.coli expression system and the effects of pH and temperature on the enzyme activity of IRED and FDH were investigated.The yield of(S)-nornicotine reached>98%with enantiomeric excess(e.e.)value of>99%under the optimal reaction conditions of temperature 30℃,pH 7.5,myosmine mass concentration 30 g/L,FDH cells 600 U/L,IRED cells 900 U/L,NADP+ 0.6 mmol/L and sodium formate mass concentration 90 g/L.Finally,(S)-nicotine with a yield and e.e.value of above 99%was obtained by chemical methylation of(S)-nornicotine.
关键词
(S)-烟碱/亚胺还原酶/甲酸脱氢酶/(S)-降烟碱/生物工程Key words
(S)-nicotine/imine reductase(IRED)/formate dehydrogenase(FDH)/(S)-nornicotine/bioengineering引用本文复制引用
基金项目
国家自然科学基金(22178083)
河北省自然科学基金(B2020202036)
河北省重点研发计划(22372802D)
出版年
2024
NETL
NSTL
维普
万方数据