酶的非天然催化功能：非水介质中酶催化的不对称Mannich反应及机理研究

Since fossil resource is continuously consuming, and environmental pollution is constantly increasing, the human have to face the unprecedented crisis of survival and development. So the traditional material processing industry must have a revolutionary change to biological processing industry which utilizes biological renewable resources as raw material and is environmentally friendly. Mannich reaction is one of the most widely used reactions, and has been used to prepare medicines, pesticides and other many chemical products by both the laboratory and enterprise. But environmentally friendly, sustainable, biocatalytic, and asymmetric methods of Mannich reaction are still lack. This project will study on the enzyme-catalyzed asymmetric Mannich reaction in non-aqueous media. (1) Found the enzymes which have asymmetric catalytic activity for Mannich reaction; (2) Study the influence of reaction conditions such as solvents, temperature, pH, water contents, metal ions and the scope and ratio of substrates on the reaction; (3) Establish the methods of high yield rate and high stereoselectivity; (4) Explore the reaction mechanism. This project aims to research and explore the regularities of enzyme-catalyzed asymmetric Mannich reaction, clarify the principle of adaptability of enzyme-catalyzed Mannich reaction in the non-natural environment, medium engineering, and controlling methods.

化石资源不断枯竭，环境污染日益加剧, 人类面临着前所未有的生存与发展危机。因此由传统的物质加工业转向以生物可再生资源为原料的、环境友好的生物加工业成为发展趋势。Mannich反应是应用最广的反应之一，用于制备医药、农药及其它许许多多化工品。开展环境友好、可持续发展的生物催化不对称Mannich反应方法研究具有重要意义。本项目拟针对应用较广的Mannich反应类型，开展非水介质中酶催化不对称反应的研究。首先遴选具有不对称催化Mannich反应活性的酶；然后研究反应条件如溶剂、温度、pH、含水量、金属离子和底物等对反应的影响及调控，建立高产率，高选择性的Mannich反应方法；进而对反应的机理进行系统研究。本项目的研究旨在阐明非水介质中酶催化Mannich反应的规律，探讨与生物催化剂相适应的人工微环境因素、介质工程及调控方法，建立可供生产需要的生物不对称催化Mannich方法。

化石资源不断枯竭，环境污染日益加剧, 人类面临着前所未有的生存与发展危机。因此由传统的物质加工业转向以生物可再生资源为原料的、环境友好的生物加工业成为发展趋势。Mannich 反应是应用最广的反应之一，用于制备医药、农药及其它许许多多化工品。开展环境友好、可持续发展的生物催化不对称Mannich 反应方法研究具有重要意义。本项目拟针对应用较广的Mannich 反应类型，开展非水介质中酶催化不对称反应的研究。I．通过比对一级和晶体结构挖掘生物催化剂；II．发展了灰色链霉菌蛋白酶 (SGP)、蜂蜜曲霉酰化酶(AMA)、蚯蚓粗提物、小麦胚脂肪酶催化的各类不对称Mannich反应，系统研究了各种影响和调控因素、反应适用范围及反应机理；III．发现了酶催化的与Mannich 反应机理类似的不对称aldol、Michael 反应、Henry 反应、Biginelli 反应、aza-Diels-Alder、Povarov反应及串联反应等；IV．拓展了可见光/酶接力催化的吲哚及衍生物的胺烷基化反应、其他绿色（无催化剂及氨基酸催化等）合成方法的研究和受邀综述水解酶催化的不对称C-C键构建的方法。