Journal
ACS SYNTHETIC BIOLOGY
Volume 9, Issue 2, Pages 304-+Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acssynbio.9b00331
Keywords
consolidated bioprocessing; butanol; Clostridium; push-pull strategy; carbon flux
Categories
Funding
- National Natural Science Foundation of China [21706133, 21825804, 31670094]
- Fundamental Research Funds for the Central Universities [30918011310]
- Key Laboratory of Biomass Chemical Engineering of Ministry of Education, Zhejiang University [2018BCE003]
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Clostridium cellulovorans DSM 743B can produce butyrate when grown on lignocellulose, but it can hardly synthesize butanol. In a previous study, C. cellulovorans was successfully engineered to switch the metabolism from butyryl-CoA to butanol by overexpressing an alcohol aldehyde dehydrogenase gene adhE1 from Clostridium acetobutylicum ATCC 824; however, its full potential in butanol production is still unexplored. In the study, a metabolic engineering approach based on a push pull strategy was developed to further enhance cellulosic butanol production. In order to accomplish this, the carbon flux from acetylCoA to butyryl-CoA was pulled by overexpressing a trans-enoyl-coenzyme A reductase gene (ter), which can irreversibly catalyze crotonyl-CoA to butyryl-CoA. Then an acid reassimilation pathway uncoupled with acetone production was introduced to redirect the carbon flow from butyrate and acetate toward butyryl-CoA. Finally, xylose metabolism engineering was implemented by inactivating xylR (Clocel_0594) and araR (Clocel_1253), as well as overexpressing xylT (CA_C1345), which is expected to supply additional carbon and reducing power for CoA and butanol synthesis pathways. The final engineered strain produced 4.96 g/L of n-butanol from alkali extracted corn cobs (AECC), increasing by 235-fold compared to that of the wild type. It serves as a promising butanol producer by consolidated bioprocessing.
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