Journal
METABOLIC ENGINEERING
Volume 20, Issue -, Pages 49-55Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymben.2013.09.003
Keywords
Redox rebalancing; Metabolic imbalance; n-butanol; Synthetic biology; Metabolic engineering
Categories
Funding
- Basic Science Research Program [2012R1A2A2A01009868]
- Advanced Biomass R & D Center of the National Research Foundation of Korea [ABC-2010-0029800]
- Ministry of Education, Science and Technology
- Gyeongbuk Sea Grant Program
- Manpower Development Program
- Marine Biotechnology Program
- Ministry of Oceans and Fisheries Korea
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The intracellular redox state plays an important role in the cellular physiology that determines the efficiency of chemical and biofuel production by microbial cell factories. However, it is difficult to achieve optimal redox rebalancing of synthetic pathways owing to the sensitive responses of cellular physiology according as the intracellular redox state changes. Here, we demonstrate optimal rebalancing of the intracellular redox state by model-driven control of expression using n-butanol production in Escherichia coli as a model system. The synthetic n-butanol production pathway was constructed by implementing synthetic constitutive promoters and designing synthetic 5'-untranslated regions (5'-UTR) for each gene. Redox rebalancing was achieved by anaerobically activating the pyruvate dehythogenase (PDH) complex and additionally tuning the expression level of NAD(+)-dependent formate dehydrogenase (fdh1 from Saccharomyces cerevisiae) through rational UTR engineering. Interestingly, efficient production of n-butanol required different amounts of reducing equivalents depending on whether the substrate was glucose or galactose. One intriguing implication of this work is that additional strain improvement can be achieved, even within given genetic components, through rebalancing intracellular redox state according to target products and substrates. (C) 2013 Elsevier Inc. All rights reserved
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