Abstract
As the consequences of climate change become apparent, metabolic engineers and synthetic biologists are exploring sustainable sources for transportation fuels. The design and engineering of microorganisms to produce bio-gasoline and other biofuels from renewable feedstocks can significantly reduce dependence on fossil fuels as well as lower the emissions of greenhouse gases. A significant amount of research over the past two decades has led to the development of microbial strains for the production of advanced fuel compounds. Crucial to these efforts are robust methods to quantify the amount of the biofuel compound being produced as well as the other metabolites that might be present during fermentation. Here, we provide a protocol for the quantification of branched-chain alcohols, specifically isobutanol and isopropanol, using high-performance liquid chromatography (HPLC).
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Liao JC, Mi L, Pontrelli S, Luo S (2016) Fuelling the future: microbial engineering for the production of sustainable biofuels. Nat Rev Microbiol 14:288–304
Keasling JD (2010) Manufacturing molecules through metabolic engineering. Science 330:1355–1358
Kung Y, Runguphan W, Keasling JD (2012) From fields to fuels: recent advances in the microbial production of biofuels. ACS Synth Biol 1:498–513. https://doi.org/10.1021/sb300074k
Peralta-Yahya PP, Zhang F, Del Cardayre SB, Keasling JD (2012) Microbial engineering for the production of advanced biofuels. Nature 488:320–328
Lan EI, Liao JC (2013) Microbial synthesis of n-butanol, isobutanol, and other higher alcohols from diverse resources. Bioresour Technol 135:339–349. https://doi.org/10.1016/j.biortech.2012.09.104
Atsumi S, Hanai T, Liao JC (2008) Non-fermentative pathways for synthesis of branched-chain higher alcohols as biofuels. Nature 451:86–89. https://doi.org/10.1038/nature06450
Avalos JL, Fink GR, Stephanopoulos G (2013) Compartmentalization of metabolic pathways in yeast mitochondria improves production of branched chain alcohols. Nat Biotechnol 31:335–341. https://doi.org/10.1038/nbt.2509.Compartmentalization
Siripong W, Wolf P, Kusumoputri TP et al (2018) Metabolic engineering of Pichia pastoris for production of isobutanol and isobutyl acetate. Biotechnol Biofuels 11:1. https://doi.org/10.1186/s13068-017-1003-x
Acknowledgments
W.R. gratefully acknowledges the support from the Thailand Research Fund under grant contract number TRG6180006.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2021 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Runguphan, W., Kocharin, K. (2021). Quantification of Branched-Chain Alcohol-Based Biofuels and Other Fermentation Metabolites via High-Performance Liquid Chromatography. In: Basu, C. (eds) Biofuels and Biodiesel. Methods in Molecular Biology, vol 2290. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1323-8_5
Download citation
DOI: https://doi.org/10.1007/978-1-0716-1323-8_5
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-1322-1
Online ISBN: 978-1-0716-1323-8
eBook Packages: Springer Protocols