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Engineered ketol-acid reductoisomerase and alcohol dehydrogenase enable anaerobic 2-methylpropan-1-ol production at theoretical yield in Escherichia coli

Bastian, Sabine and Liu, Xiang and Meyerowitz, Joseph T. and Snow, Christopher D. and Chen, Mike M. Y. and Arnold, Frances H. (2011) Engineered ketol-acid reductoisomerase and alcohol dehydrogenase enable anaerobic 2-methylpropan-1-ol production at theoretical yield in Escherichia coli. Metabolic Engineering, 13 (3). pp. 345-352. ISSN 1096-7176. doi:10.1016/j.ymben.2011.02.004. https://resolver.caltech.edu/CaltechAUTHORS:20110526-140043166

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Abstract

2-methylpropan-1-ol (isobutanol) is a leading candidate biofuel for the replacement or supplementation of current fossil fuels. Recent work has demonstrated glucose to isobutanol conversion through a modified amino acid pathway in a recombinant organism. Although anaerobic conditions are required for an economically competitive process, only aerobic isobutanol production has been feasible due to an imbalance in cofactor utilization. Two of the pathway enzymes, ketol-acid reductoisomerase and alcohol dehydrogenase, require nicotinamide dinucleotide phosphate (NADPH); glycolysis, however, produces only nicotinamide dinucleotide (NADH). Here, we compare two solutions to this imbalance problem: (1) over-expression of pyridine nucleotide transhydrogenase PntAB and (2) construction of an NADH-dependent pathway, using engineered enzymes. We demonstrate that an NADH-dependent pathway enables anaerobic isobutanol production at 100% theoretical yield and at higher titer and productivity than both the NADPH-dependent pathway and transhydrogenase over-expressing strain. Our results show how engineering cofactor dependence can overcome a critical obstacle to next-generation biofuel commercialization.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1016/j.ymben.2011.02.004 DOIArticle
http://www.sciencedirect.com/science/article/pii/S1096717611000164PublisherArticle
ORCID:
AuthorORCID
Snow, Christopher D.0000-0002-7690-3519
Arnold, Frances H.0000-0002-4027-364X
Additional Information:© 2011 Elsevier Inc. Available online 29 April 2011. Research was sponsored by the U.S. Army Research Laboratory and was accomplished under cooperative Agreement no. W911NF-09-2-0022. The views and conclusions contained in this document are those of the authors and should not be interpreted as representing the official policies, either expressed or implied, of the Army Research Laboratory or the U.S. Government. The U.S. Government is authorized to reproduce and distribute reprints for Government purposes notwithstanding any copyright notation herein. J.T.M. is supported by the Department of Defense through the National Defense Science & Engineering Graduate Fellowship Program and by the National Science Foundation through the Graduate Research Fellowship Program. X.L. is supported by the China Scholarship Council (CSC).
Funders:
Funding AgencyGrant Number
Army Research LaboratoryW911NF-09-2-0022
National Defense Science and Engineering Graduate (NDSEG) FellowshipUNSPECIFIED
NSF Graduate Research FellowshipUNSPECIFIED
China Scholarship CouncilUNSPECIFIED
Subject Keywords:Metabolic engineering; Cofactor imbalance; Isobutanol; Ketol-acid reductoisomerase; Biofuels
Issue or Number:3
DOI:10.1016/j.ymben.2011.02.004
Record Number:CaltechAUTHORS:20110526-140043166
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20110526-140043166
Official Citation:Sabine Bastian, Xiang Liu, Joseph T. Meyerowitz, Christopher D. Snow, Mike M.Y. Chen, Frances H. Arnold, Engineered ketol-acid reductoisomerase and alcohol dehydrogenase enable anaerobic 2-methylpropan-1-ol production at theoretical yield in Escherichia coli, Metabolic Engineering, Volume 13, Issue 3, May 2011, Pages 345-352, ISSN 1096-7176, DOI: 10.1016/j.ymben.2011.02.004. (http://www.sciencedirect.com/science/article/pii/S1096717611000164)
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:23812
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:01 Jun 2011 18:20
Last Modified:09 Nov 2021 16:18

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