体外无细胞紫杉醇及其前驱体生物合成系统的构建

项目名称： 体外无细胞紫杉醇及其前驱体生物合成系统的构建

项目编号： No.21476167

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 有机化学

项目作者： 齐浩

作者单位： 天津大学

项目金额： 84万元

中文摘要： 针对复杂代谢产物如紫杉醇生物合成困难问题,申请者提出了开发介于死的化学合成与活的生物合成之间的体外代谢合成系统, 结合先进的人工DNA折叠纳米结构技术，开发具有精密空间可控性的体外串联代谢分子机器。主要研究内容为：1）在体外构建紫杉醇合成中包括基于P450酶的，IPP聚合、母核成环以及下游的羟化、酰基转移反应的生物合成系统，2）通过设计DNA折叠纳米结构体，精密地控制代谢酶的空间位置，实现串联代谢分子机器。该研究思路和研究方案具有明显创新性，本研究可望成为体外合成生物系统的一个经典案例，为系统生物学的研究提供全新的技术，为实现产业化提供基础。还可以加深对紫杉醇的生物合成途径反应机理理解，成为减少生物工程中对细胞过渡依赖的崭新技术，可预计此技术可转化应用到更多的生物工程研究中。

中文关键词： 生物合成；生物化工；过程优化

英文摘要： With the development of synthetic biotechnology on genome and protein enzyme engineering, re-designed metabolism reaction networks showed great potential for mass manufacturing chemical with complicated structures, which is very difficult to be synthesized commercially using traditional chemical method. Facing the more and more serous environmental problem, reengineering bio-metabolism brougt us an alternative way for synthesis of material of high value. Although, many progressed has been made in engineering cell, such as E.coli and yeast, for biosynthesis of chemcial compound in the past few years. However, it is still very challenging to tame the cell for artifical purposes, because compolicated endogenic metabolism network for maintenaning cell. Here, we propose a new strategy in which a complete new cell free metabolism network will be constructed from purified metabolism enzymes for chemical synthesis in test tube. Furthermore, DNA origami will be used to regulate spacial position of the metabolims enzymes in vitro. Generally, one metabolism network was comprised from many enzymes and chemical was synthesized through multiple steps in a series circuits. For mimicking and regulating the reaction flow in the in vitro system, enzymes will be imobilized on the designed DNA scaffold in the order, by which they get involed in the metabolims flow. It will be a completed new artifical complicate biomolecular machine, which we called as metabolism robotic. In this proposal, we will focus on P450 enzyme system related part in Taxol metablosim, which is the bottleneck restricting mass biosynthesis of taxol in engineered cell. There is lots of advantage in this new technology in comparison with in vivo metabolism engineering. Because only components that are necessary for the target chemical synthesis are included, there should no interference with other bioreaction that happened in the cell in vivo. Similarly, it should be easy to connect metabolism reaction module from different cell sources. Therefore, the in vitro system will be a flexiable platform for various bioengineering purposes.

英文关键词： biosynthesis;Biochemistry;bioreaction optimization