Improving d-glucaric acid production from myo-inositol in E. coli by increasing MIOX stability and myo-inositol transport
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MBE-13-181R1 extracted pages.pdf
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2.03 MB
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Author(s)
Shiue, Eric
Prather, Kristala L
Date Issued
December 2013
Journal
Metabolic Engineering
Publisher
Elsevier
Citation
Shiue, Eric, and Kristala L.J. Prather. “Improving d-Glucaric Acid Production from Myo-Inositol in E. Coli by Increasing MIOX Stability and Myo-Inositol Transport.” Metabolic Engineering 22 (March 2014): 22–31.
Version
Author's final manuscript
Abstract
d-glucaric acid has been explored for a myriad of potential uses, including biopolymer production and cancer treatment. A biosynthetic route to produce d-glucaric acid from glucose has been constructed in Escherichia coli ( Moon et al., 2009b), and analysis of the pathway revealed myo-inositol oxygenase (MIOX) to be the least active enzyme. To increase pathway productivity, we explored protein fusion tags for increased MIOX solubility and directed evolution for increased MIOX activity. An N-terminal SUMO fusion to MIOX resulted in a 75% increase in d-glucaric acid production from myo-inositol. While our directed evolution efforts did not yield an improved MIOX variant, our screen isolated a 941 bp DNA fragment whose expression led to increased myo-inositol transport and a 65% increase in d-glucaric acid production from myo-inositol. Overall, we report the production of up to 4.85 g/L of d-glucaric acid from 10.8 g/L myo-inositol in recombinant E. coli.
MIT Department
Massachusetts Institute of Technology. Department of Chemical Engineering
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Creative Commons Attribution-NonCommercial-NoDerivs License
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DOI of Published Version
https://doi.org/10.1016/j.ymben.2013.12.002
