Abstract
Clostridium acetobutylicum is a bacterial species that ferments sugar to a mixture of organic solvents (acetone, butanol and ethanol). This protocol delineates a methodology to combine solventogenic clostridial fermentation and chemical catalysis via extractive fermentation for the production of biofuel blendstocks. Extractive fermentation of C. acetobutylicum is operated in fed-batch mode with a concentrated feed solution (500 grams per liter glucose and 50 grams per liter yeast extract) for 60 h, producing in excess of 40 g of solvents (acetone, butanol and ethanol) between the completely immiscible extractant and aqueous phases of the bioreactor. After distillation of the extractant phase, the acetone, butanol and ethanol mixture is upgraded to long-chain ketones over a palladium-hydrotalcite (Pd-HT) catalyst. This reaction is generally carried out in batch with a high-pressure Q-tube for 20 h at 250 °C. Following this protocol enables the production of ∼0.5 g of high-value biofuel precursors from a 1.7-g portion of fermentation solvents.
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Acknowledgements
This work was funded by the Energy Biosciences Institute.
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S.S. and Z.C.B. contributed equally to this work. S.S., P.A., G.G. and A.G. performed experiments on the chemical catalysts, Z.C.B. optimized the fermentation and extractive processes. All authors contributed to the conception of the experiments, discussion of the results and preparation of the manuscript.
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Sreekumar, S., Baer, Z., Pazhamalai, A. et al. Production of an acetone-butanol-ethanol mixture from Clostridium acetobutylicum and its conversion to high-value biofuels. Nat Protoc 10, 528–537 (2015). https://doi.org/10.1038/nprot.2015.029
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DOI: https://doi.org/10.1038/nprot.2015.029
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