title: In Vitro Reconstitution and Optimization of the Entire Pathway to Convert Glucose into Fatty Acid.
abstract: 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.
authors.affiliation: Department of Endocrinology, Zhongnan Hospital of Wuhan University , Wuhan 430071, China.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education and Wuhan University School of Pharmaceutical Sciences , Wuhan 430071, China.
authors.forename: Yuchen
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education and Wuhan University School of Pharmaceutical Sciences , Wuhan 430071, China.
authors.forename: Xiaoge
authors.initials: X
authors.lastname: Jia
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education and Wuhan University School of Pharmaceutical Sciences , Wuhan 430071, China.
authors.forename: Mengzhu
authors.initials: M
authors.lastname: Hu
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education and Wuhan University School of Pharmaceutical Sciences , Wuhan 430071, China.
authors.forename: Zixin
authors.initials: Z
authors.lastname: Deng
authors.affiliation: Department of Endocrinology, Zhongnan Hospital of Wuhan University , Wuhan 430071, China.
authors.forename: Yancheng
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education and Wuhan University School of Pharmaceutical Sciences , Wuhan 430071, China.
authors.forename: Tiangang
authors.identifier: 0000-0001-8087-0345
authors.initials: T
authors.lastname: Liu
mesh_terms: D000103:Acetyl-CoA Carboxylase / Q000235:genetics / Q000378:metabolism; D002855:Chromatography, Thin Layer; D004926:Escherichia coli / Q000378:metabolism*; D064429:Fatty Acid Synthases / Q000235:genetics / Q000378:metabolism; D005227:Fatty Acids / Q000032:analysis / Q000378:metabolism*; D008401:Gas Chromatography-Mass Spectrometry; D005947:Glucose / Q000378:metabolism*; D006019:Glycolysis; D009249:NADP / Q000378:metabolism; D010427:Pentose Phosphate Pathway; D011768:Pyruvate Dehydrogenase Complex / Q000235:genetics / Q000378:metabolism
chemical_list: D005227:Fatty Acids; D011768:Pyruvate Dehydrogenase Complex; D009249:NADP; D064429:Fatty Acid Synthases; D000103:Acetyl-CoA Carboxylase; D005947:Glucose
keywords: fatty acid biosynthesis; fatty acids; glucose; glycolysis; in vitro reconstitution; pyruvate dehydrogenase