Metabolic engineering of Escherichia coli for direct production of 1,4-butanediol

• Published in: Nature chemical biology Vol. 7; no. 7; pp. 445 - 452
• Main Authors: Yim, Harry, Haselbeck, Robert, Niu, Wei, Pujol-Baxley, Catherine, Burgard, Anthony, Boldt, Jeff, Khandurina, Julia, Trawick, John D, Osterhout, Robin E, Stephen, Rosary, Estadilla, Jazell, Teisan, Sy, Schreyer, H Brett, Andrae, Stefan, Yang, Tae Hoon, Lee, Sang Yup, Burk, Mark J, Van Dien, Stephen
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 22.05.2011; Nature Publishing Group
• Subjects: 631/326/252/318; 631/92/60; 631/92/603; Algorithms; Anaerobiosis; Biochemical Engineering; Biochemistry; Bioengineering; Bioorganic Chemistry; Biosynthesis; Biosynthetic Pathways; Butylene Glycols - chemistry; Butylene Glycols - metabolism; Cell Biology; Chemicals; Chemistry; Chemistry and Materials Science; Chemistry/Food Science; E coli; Escherichia coli - enzymology; Escherichia coli - genetics; Escherichia coli - metabolism; Fermentation; Genetic Engineering; Glucose - metabolism; Metabolism; Natural gas; Organisms, Genetically Modified - metabolism; Polymers; Sugar
• ISSN: 1552-4450; 1552-4469
• DOI: 10.1038/nchembio.580

The design and implementation of a high-yielding enzymatic route to 1,4-butanediol—a compound not known to be produced naturally—provides a compelling example of how metabolic engineering can be harnessed for the microbial conversion of carbohydrate feedstocks to desired small molecules. 1,4-Butanediol (BDO) is an important commodity chemical used to manufacture over 2.5 million tons annually of valuable polymers, and it is currently produced exclusively through feedstocks derived from oil and natural gas. Herein we report what are to our knowledge the first direct biocatalytic routes to BDO from renewable carbohydrate feedstocks, leading to a strain of Escherichia coli capable of producing 18 g l −1 of this highly reduced, non-natural chemical. A pathway-identification algorithm elucidated multiple pathways for the biosynthesis of BDO from common metabolic intermediates. Guided by a genome-scale metabolic model, we engineered the E. coli host to enhance anaerobic operation of the oxidative tricarboxylic acid cycle, thereby generating reducing power to drive the BDO pathway. The organism produced BDO from glucose, xylose, sucrose and biomass-derived mixed sugar streams. This work demonstrates a systems-based metabolic engineering approach to strain design and development that can enable new bioprocesses for commodity chemicals that are not naturally produced by living cells.