Journal
CELL SYSTEMS
Volume 3, Issue 2, Pages 133-143Publisher
CELL PRESS
DOI: 10.1016/j.cels.2016.07.012
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Funding
- Competitive Research Programme of the National Research Foundation of Singapore [NRF-CRP5-2009-03]
- Agency for Science, Technology, and Research of Singapore [1324004108]
- Synthetic Biology Initiative of the National University of Singapore [DPRT/943/09/14]
- Defense Threat Reduction Agency [HDTRA1-13-1-0037]
- Energy Innovation Programme Office of the Economic Development Board of Singapore
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We present a synthetic gene circuit for decoupling cell growth from metabolite production through autonomous regulation of enzymatic pathways by integrated modules that sense nutrient and substrate. The two-layer circuit allows Escherichia coli to selectively utilize target substrates in a mixed pool; channel metabolic resources to growth by delaying enzymatic conversion until nutrient depletion; and activate, terminate, and re-activate conversion upon substrate availability. We developed two versions of controller, both of which have glucose nutrient sensors but differ in their substrate-sensing modules. One controller is specific for hydroxycinnamic acid and the other for oleic acid. Our hydroxycinnamic acid controller lowered metabolic stress 2-fold and increased the growth rate 2-fold and productivity 5-fold, whereas our oleic acid controller lowered metabolic stress 2-fold and increased the growth rate 1.3-fold and productivity 2.4-fold. These results demonstrate the potential for engineering strategies that decouple growth and production to make bio-based production more economical and sustainable.
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