Journal
METABOLIC ENGINEERING
Volume 38, Issue -, Pages 389-400Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymben.2016.10.002
Keywords
Gas fermentation; Clostridium ljungdahlii; In silico metabolic engineering; OptKnock; Spatiotemporal metabolic modeling
Categories
Funding
- National Science Foundation [CBET 1511346]
- Div Of Chem, Bioeng, Env, & Transp Sys
- Directorate For Engineering [1511346] Funding Source: National Science Foundation
- The British Council [GII201] Funding Source: researchfish
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Synthesis gas fermentation is one of the most promising routes to convert synthesis gas (syngas; mainly comprised of H-2 and CO) to renewable liquid fuels and chemicals by specialized bacteria. The most commonly studied syngas fermenting bacterium is Clostridium ljungdahlii, which produces acetate and ethanol as its primary metabolic byproducts. Engineering of C. ljungdahlii metabolism to overproduce ethanol, enhance the synthesize of the native byproducts lactate and 2,3-butanediol, and introduce the synthesis of non-native products such as butanol and butyrate has substantial commercial value. We performed in silico metabolic engineering studies using a genome-scale reconstruction of C. ljungdahlii metabolism and the OptKnock computational framework to identify gene knockouts that were predicted to enhance the synthesis of these native products and non-native products, introduced through insertion of the necessary heterologous pathways. The OptKnock derived strategies were often difficult to assess because increase product synthesis was invariably accompanied by decreased growth. Therefore, the OptKnock strategies were further evaluated using a spatiotemporal metabolic model of a syngas bubble column reactor, a popular technology for large-scale gas fermentation. Unlike flux balance analysis, the bubble column model accounted for the complex tradeoffs between increased product synthesis and reduced growth rates of engineered mutants within the spatially varying column environment. The two-stage methodology for deriving and evaluating metabolic engineering strategies was shown to yield new C. ljungdahlii gene targets that offer the potential for increased product synthesis under realistic syngas fermentation conditions.
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