R,R-2,3-丁二醇是一种新兴的微生物四碳手性醇，超高光学纯度（≥99.9%）的该产品是合成手性电子材料、医药中间体的重要原料，近年来，受到了化工、材料界的广泛关注。本课题针对Paenibacillus polymyxa发酵制备R,R-2,3-丁二醇过程中存在少量光学副产物meso-2,3-丁二醇以及NADH辅酶再生问题，巧妙地运用基因替换策略，构建新型λRed 同源重组系统，将该菌中丁二酮还原酶基因替换为NAD+依赖型甲酸脱氢酶基因，在阻断meso-2,3-丁二醇合成的同时偶联NADH辅酶再生，并借助最佳代谢流优化方案的设计，对辅酶、碳平衡进行有效调控，最终实现超高光学纯R,R-2,3-丁二醇的高效制备，促进其生产技术的发展以及在手性产品合成中的应用，并为类似手性醇、手性氨基酸等手性有机小分子的研究提供借鉴，具有重要的学术意义和实用价值。

R,R-2,3-butanediol, which is a kind of microbial production of four carbon chiral alcohol, has attracted increasing interest of researchers in the chemical and material engineering field because of its diverse industrial applications in the chiral synthesis in recent years. The R,R-2,3-butanediol at higher than 99.9% optical purity has practical uses in the synthesis of some chiral electronic material and chiral pharmaceuticals. In this research, the new method about the genetic manipulation of R,R-2,3-butanediol metabolic pathway in Paenibacillus polymyxa will be studied in order to reduce the small by-product meso-2,3-butanediol and achieve cefactor NADH regeneration. The gene of diacetyl reductase in P. polymyxa will be skillfully replaced with the gene of NAD-dependent formate dehydrogenase using the λ Red homologous recombination technique, and the synthesis of meso-2,3-butanediol will be interrupted as the same time the coenzyme NADH is regenerated. Then, the metabolic flux in the recombinant strain are efficiently regulated to optimize the coenzyme and carbon balance. Finally,the R,R-2,3-butanediol at higher than 99.9% optical purity and at higher yield will be achieved success. This work will promote the technological development of industrial R,R-2,3-butanediol production and its application in the chiral synthesis, and provide a kind of method, which will be utilized in the study of the chiral small molecule organic compounds, such as other chiral alcohol, chiral amino acid and so on. Through above-mentioned research, the huge economic and social benefit could been obtained.