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Uncovering rare NADH-preferring ketol-acid reductoisomerases

Brinkmann-Chen, S. and Cahn, J. K. B. and Arnold, Frances H. (2014) Uncovering rare NADH-preferring ketol-acid reductoisomerases. Metabolic Engineering, 26 . pp. 17-22. ISSN 1096-7176. http://resolver.caltech.edu/CaltechAUTHORS:20140902-115405020

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Abstract

All members of the ketol-acid reductoisomerase (KARI) enzyme family characterized to date have been shown to prefer the nicotinamide adenine dinucleotide phosphate hydride (NADPH) cofactor to nicotinamide adenine dinucleotide hydride (NADH). However, KARIs with the reversed cofactor preference are desirable for industrial applications, including anaerobic fermentation to produce branched-chain amino acids. By applying insights gained from structural and engineering studies of this enzyme family to a comprehensive multiple sequence alignment of KARIs, we identified putative NADH-utilizing KARIs and characterized eight whose catalytic efficiencies using NADH were equal to or greater than NADPH. These are the first naturally NADH-preferring KARIs reported and demonstrate that this property has evolved independently multiple times, using strategies unlike those used previously in the laboratory to engineer a KARI cofactor switch.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1016/j.ymben.2014.08.003DOIArticle
http://www.sciencedirect.com/science/article/pii/S1096717614001062#PublisherArticle
http://www.sciencedirect.com/science/MiamiMultiMediaURL/1-s2.0-S1096717614001062/1-s2.0-S1096717614001062-mmc1.doc/272595/FULL/S1096717614001062/01de2dfd04baa99e883ee8d7af673b64/mmc1.docPublisherSupplemental Material
http://www.sciencedirect.com/science/MiamiMultiMediaURL/1-s2.0-S1096717614001062/1-s2.0-S1096717614001062-mmc2.doc/272595/FULL/S1096717614001062/0c24d01304969f1aa32261003f4b8912/mmc2.docPublisherSupplemental Material
ORCID:
AuthorORCID
Arnold, Frances H.0000-0002-4027-364X
Additional Information:© 2014 Elsevier B.V. Received 20 June 2014, Revised 1 August 2014, Accepted 19 August 2014, Available online 27 August 2014. J.K.B.C. acknowledges the support of the Resnick Sustainability Institute (Caltech). This publication is funded by the Gordon and Betty Moore Foundation through Grant GBMF2809 to the Caltech Programmable Molecular Technology Initiative. Author contributions: SBC, JKBC, and FHA designed research. SBC and JKBC performed research and analyzed data. SBC, JKBC, and FHA wrote the paper.
Group:Resnick Sustainability Institute
Funders:
Funding AgencyGrant Number
Resnick Sustainability InstituteUNSPECIFIED
Gordon and Betty Moore FoundationGBMF2809
Caltech Programmable Molecular Technology InitiativeUNSPECIFIED
Non-Subject Keywords:Ketol acid reductoisomerase; Cofactor specificity; NADH; NADPH
Record Number:CaltechAUTHORS:20140902-115405020
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20140902-115405020
Official Citation:S. Brinkmann-Chen, J.K.B. Cahn, F.H. Arnold, Uncovering rare NADH-preferring ketol-acid reductoisomerases, Metabolic Engineering, Volume 26, November 2014, Pages 17-22, ISSN 1096-7176, http://dx.doi.org/10.1016/j.ymben.2014.08.003. (http://www.sciencedirect.com/science/article/pii/S1096717614001062)
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:49116
Collection:CaltechAUTHORS
Deposited By: SWORD User
Deposited On:02 Sep 2014 20:29
Last Modified:08 Sep 2017 23:00

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