National Energy R&D Center for Biorefinery, Beijing Key Laboratory of Bioprocess, Beijing University of Chemical Technology, 15th, Beisanhuan East Road, Beijing, 100029, PR China.
Biotechnol J. 2017 Nov;12(11). doi: 10.1002/biot.201700176. Epub 2017 Sep 6.
Diols, such as 1,3-propanediol (1,3-PDO) and 2,3-butanediol (2,3-BDO), have several promising properties for many synthetic reactions. Here, the cofactor engineering strategy, including the construction of Entner-Doudoroff pathway and transhydrogenase-based NADH regeneration system, was applied in producing diols from mixtures of glucose and glycerol. Entner-Doudoroff pathway had a high regeneration rate of NAD(P)H. This work described a strategy to administrate intracellular NADH/NAD ratio and improved the concentration of diols. The improvement of NADH/NAD ratio also effected gene transcription level of the central carbon pathway and cell growth. Finally, the intracellular NADH/NAD ratio in KP-APZDUT was increased by 92.8% compared to the KP-T and the concentration, yield and productivity of diols were increased to 110.8 g L , 0.78 mol mol , and 3.46 g Lh , respectively. The strategy described here provides an approach to achieve a recombinant strain which is capable of producing diols with high yield and productivity. To the best of our knowledge, the Entner-Doudoroff pathway has not yet been used to produce 1,3-PDO or 2,3-BDO in Klebsiella pneumoniae.
二醇，如 1,3-丙二醇（1,3-PDO）和 2,3-丁二醇（2,3-BDO），在许多合成反应中具有多种有前途的性质。在这里，辅酶工程策略，包括构建 Entner-Doudoroff 途径和基于转氢酶的 NADH 再生系统，被应用于从葡萄糖和甘油的混合物中生产二醇。Entner-Doudoroff 途径具有很高的 NAD(P)H 再生速率。这项工作描述了一种调节细胞内 NADH/NAD 比并提高二醇浓度的策略。NADH/NAD 比的提高还影响了中心碳途径的基因转录水平和细胞生长。最后，与 KP-T 相比，KP-APZDUT 中的细胞内 NADH/NAD 比增加了 92.8%，二醇的浓度、产率和生产效率分别提高到 110.8 g/L、0.78 mol/mol 和 3.46 g/Lh。这里描述的策略为实现能够以高产率和高生产效率生产二醇的重组菌株提供了一种方法。据我们所知，Entner-Doudoroff 途径尚未在肺炎克雷伯氏菌中用于生产 1,3-PDO 或 2,3-BDO。
