Escherichia coli Deletion Mutants Illuminate Trade-Offs between Growth Rate and Flux through a Foreign Anabolic Pathway
Kelly C Falls,
Aimee L Williams,
Anton V Bryksin and
Ichiro Matsumura
PLOS ONE, 2014, vol. 9, issue 2, 1-8
Abstract:
Metabolic engineers strive to improve the production yields of microbial fermentations, sometimes by mutating the genomes of production strains. Some mutations are detrimental to the health of the organism, so a quantitative and mechanistic understanding of the trade-offs could inform better designs. We employed the bacterial luciferase operon (luxABCDE), which uses ubiquitous energetic cofactors (NADPH, ATP, FMNH2, acetyl-CoA) from the host cell, as a proxy for a novel anabolic pathway. The strains in the Escherichia coli Keio collection, each of which contains a single deletion of a non-essential gene, represent mutational choices that an engineer might make to optimize fermentation yields. The Keio strains and the parental BW25113 strain were transformed with a luxABCDE expression vector. Each transformant was propagated in defined M9 medium at 37°C for 48 hours; the cell density (optical density at 600 nanometers, OD600) and luminescence were measured every 30 minutes. The trade-offs were visualized by plotting the maximum growth rate and luminescence/OD600 of each transformant across a “production possibility frontier”. Our results show that some loss-of-function mutations enhance growth in vitro or light production, but that improvement in one trait generally comes at the expense of the other.
Date: 2014
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Persistent link: https://EconPapers.repec.org/RePEc:plo:pone00:0088159
DOI: 10.1371/journal.pone.0088159
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