为实现甲醇等一碳资源的有效利用，通过合成生物学技术人工设计构建新的一碳代谢途径是近年来的研究热点。羟基乙醛合成酶（GALS）是人工设计的能够催化甲醛生成羟基乙醛的一类甲醛缩合酶，能够实现一碳单位到二碳的线性转化，在人工一碳途径设计中受到广泛关注。本项目针对GALS催化活性低、无法满足细胞内代谢通量需求的问题，开发迭代定向进化系统结合底盘设计优化GALS的催化活性。进而通过13C代谢通量分析，转录组分析并结合逆向代谢工程策略解析和优化GALS在细胞代谢网络中的适配性。本项目的实施对进一步解析GALS的催化机理，深入挖掘GALS在甲醇代谢途径设计和菌株构建中的潜力，及最终实现甲醇等一碳化合物的生物转化有重要研究意义。

In order to realize the effective utilization of one-carbon compounds such as methanol, it has become a hot topic in recent years that to design and construct a new one-carbon metabolic pathway by synthetic biological methods. Glycolaldehyde synthase is a type of condensing enzyme that is artificially designed to catalyze formaldehyde to form glycolaldehyde. It can realize the linear conversion of one carbon unit to two carbons, and has been widely concerned in the design of artificial one carbon pathway. In order to solve the problem of low catalytic activity and insufficient intracellular metabolic flux of GALS, this project developed an iterative directed evolution system to optimize the catalytic activity of GALS. Furthermore, in order to analyze and optimize the adaptability of GALS in the methanol metabolic pathway and cell metabolism network, 13C metabolic flux analysis, transcriptome analysis and reverse metabolic engineering regulation strategy will be implemented in this project. Finally, the catalytic mechanism of GALS will be further analyzed, and its potential in the methanol metabolic pathway and engineering strain construction will be further explored. It has great research significance for biotransformation of one-carbon compounds such as methanol.