基于合成生物学的丹参酚酸类成分基因尺度代谢网络调控机制研究

It is difficult to improve specific compound content greatly and precisely, this project aims at series of problems in medicinal model plant Salvia miltiorrhiza like low content of important Salvianolic acid, in addition, traditional genetic engineering method is workload, low efficiency and strong randomicity. Project also plans to draw the genome wide metabolic network model of salvianic acid content. We attend to establish a Salvianolic acid biosynthesis directional gene knockout mutant library, to analysis transcriptional groups and metabolic groups and provide direct experimental evidence on gene function of metabolic pathway structure. By the project, we can analysis gene-to-gene, gene-to-compound distribution in various metabolic network nodes and transmission and feedback regulation mechanism. Along with Simulate metabolic network, establish mathematical model and predict optimization target sides. We comprehensively annotate gene function of Salvianolic acid regulation approaches on gene level, achieve precise regulation of the Salvianolic acid biosynthesis and maximum production efficiency on the genome scale. This project will deeply explore the potential of plant secondary metabolites production, greatly expand the scope of biosynthesis in the field of natural product research and significantly influence on innovative research of medicinal plants in China.

本项目针对以药用模式植物丹参中丹酚酸类成分为代表的系列重要化合物在生物体内含量低，用传统的基因工程方法调控效率低，随机性强，难以大幅度提高特定化合物产量的问题，以绘制丹酚酸类成分基因组尺度代谢网络模型为目标。拟开展创制丹参酚酸类成分生物合成途径定向敲除突变体库，进行转录组和代谢组分析，提供代谢途径结构基因功能的直接实验证据，分析各代谢网络节点中基因-基因之间，基因-化合物之间的分配、传递和反馈等调控机制，模拟代谢网络，建立数学模型，预测优化目标靶点。通过使用合成生物学方法，在基因水平上全面注释丹酚酸调控途径基因功能，实现基因组尺度上对丹酚酸类物质生物合成精准调控和生产效率最大化。通过本项目的研究，深度开发植物生产次生代谢产物的潜力，极大的拓展了生物合成学在天然产物研究领域的范围，对我国药用植物的创新性研究具有重大意义。

本项目针对以药用模式植物丹参，使用合成生物学研究方法，联合转录因子工程学原理，创制了丹参次生代谢产物生物合成途径定向敲除突变体，进行转录组和代谢组分析，提供代谢途径结构基因和转录因子功能的直接实验证据，分析其调控代谢网络节点中基因-基因之间，发现了5个含量较低，且具有潜在成药性的化合物与基因之间的调控关系。为下一步深入研究奠定了良好的基础。通过本项目的研究，对植物生产低含量次生代谢产物的调控做出了较好的探索，拓展了生物合成学在天然产物研究领域的范围，对我国药用植物的创新性研究具有借鉴意义。