Liu Zheng, Zhang Yuchen, Jia Xiaoge, Hu Mengzhu, Deng Zixin, Xu Yancheng, Liu Tiangang
Department of Endocrinology, Zhongnan Hospital of Wuhan University , Wuhan 430071, China.
Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education and Wuhan University School of Pharmaceutical Sciences , Wuhan 430071, China.
ACS Synth Biol. 2017 Apr 21;6(4):701-709. doi: 10.1021/acssynbio.6b00348. Epub 2017 Jan 24.
Glucose and fatty acids play essential physiological roles in nearly all living organisms, and the pathway that converts glucose into fatty acid is pivotal to the central metabolic network. We have successfully reconstituted a pathway that converts glucose to fatty acid in vitro using 30 purified proteins. Through systematic titration and optimization of the glycolytic pathway and pyruvate dehydrogenase, we increased the yield of free fatty acid from nondetectable to a level that exceeded 9% of the theoretical yield. We also reconstituted the entire pentose-phosphate pathway of Escherichia coli and established a pentose phosphate-glycolysis hybrid pathway, replacing GAPDH to enhance NADPH availability. Our efforts provide a useful platform for research involving these core biochemical transformations.
葡萄糖和脂肪酸在几乎所有生物中都发挥着重要的生理作用，而将葡萄糖转化为脂肪酸的途径对于中心代谢网络至关重要。我们已成功在体外利用30种纯化蛋白重建了一条将葡萄糖转化为脂肪酸的途径。通过对糖酵解途径和丙酮酸脱氢酶进行系统滴定和优化，我们将游离脂肪酸的产量从检测不到提高到超过理论产量9%的水平。我们还重建了大肠杆菌的整个磷酸戊糖途径，并建立了一条磷酸戊糖-糖酵解杂交途径，取代甘油醛-3-磷酸脱氢酶以提高烟酰胺腺嘌呤二核苷酸磷酸（NADPH）的可用性。我们的工作为涉及这些核心生化转化的研究提供了一个有用的平台。
