Growth-coupled bioconversion of levulinic acid to butanone
Abstract
Common strategies for conversion of lignocellulosic biomass to chemical products center on deconstructing biomass polymers into fermentable sugars. Here, we demonstrate an alternative strategy, a growth-coupled, high-yield bioconversion, by feeding cells a non-sugar substrate, by-passing central metabolism, and linking a key metabolic step to generation of acetyl-CoA that is required for biomass and energy generation. Specifically, we converted levulinic acid (LA), an established degradation product of lignocellulosic biomass, to butanone (a.k.a. methyl-ethyl ketone - MEK), a widely used industrial solvent. Our strategy combines a catabolic pathway from Pseudomonas putida that enables conversion of LA to 3-ketovaleryl-CoA, a CoA transferase that generates 3-ketovalerate and acetyl-CoA, and a decarboxylase that generates 2-butanone. By removing the ability of E. coli to consume LA and supplying excess acetate as a carbon source, we built a strain of E. coli that could convert LA to butanone at high yields, but at the cost of significant acetate consumption. Using flux balance analysis as a guide, we built a strain of E. coli that linked acetate assimilation to production of butanone. This strain was capable of complete bioconversion of LA to butanone with a reduced acetate requirement and increased specific productivity. To demonstrate the bioconversion on realmore »
- Authors:
-
- Univ. of Wisconsin-Madison, Madison, WI (United States)
- Publication Date:
- Research Org.:
- Univ. of Wisconsin-Madison, Madison, WI (United States). Great Lakes Bioenergy Research Center
- Sponsoring Org.:
- USDOE Office of Science (SC), Biological and Environmental Research (BER)
- OSTI Identifier:
- 1573019
- Alternate Identifier(s):
- OSTI ID: 1630367
- Grant/Contract Number:
- SC0018409; FC02-07ER64494
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Metabolic Engineering
- Additional Journal Information:
- Journal Volume: 55; Journal Issue: C; Journal ID: ISSN 1096-7176
- Publisher:
- Elsevier
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; Bioconversion; Levulinic acid; Sustainability; Green chemistry; Metabolic engineering; Escherichia coli; Acetate; Growth-coupling; Flux balance analysis
Citation Formats
Mehrer, Christopher R., Rand, Jacqueline M., Incha, Matthew R., Cook, Taylor B., Demir, Benginur, Motagamwala, Ali Hussain, Kim, Daniel, Dumesic, James A., and Pfleger, Brian F. Growth-coupled bioconversion of levulinic acid to butanone. United States: N. p., 2019.
Web. doi:10.1016/j.ymben.2019.06.003.
Mehrer, Christopher R., Rand, Jacqueline M., Incha, Matthew R., Cook, Taylor B., Demir, Benginur, Motagamwala, Ali Hussain, Kim, Daniel, Dumesic, James A., & Pfleger, Brian F. Growth-coupled bioconversion of levulinic acid to butanone. United States. https://doi.org/10.1016/j.ymben.2019.06.003
Mehrer, Christopher R., Rand, Jacqueline M., Incha, Matthew R., Cook, Taylor B., Demir, Benginur, Motagamwala, Ali Hussain, Kim, Daniel, Dumesic, James A., and Pfleger, Brian F. Wed .
"Growth-coupled bioconversion of levulinic acid to butanone". United States. https://doi.org/10.1016/j.ymben.2019.06.003. https://www.osti.gov/servlets/purl/1573019.
@article{osti_1573019,
title = {Growth-coupled bioconversion of levulinic acid to butanone},
author = {Mehrer, Christopher R. and Rand, Jacqueline M. and Incha, Matthew R. and Cook, Taylor B. and Demir, Benginur and Motagamwala, Ali Hussain and Kim, Daniel and Dumesic, James A. and Pfleger, Brian F.},
abstractNote = {Common strategies for conversion of lignocellulosic biomass to chemical products center on deconstructing biomass polymers into fermentable sugars. Here, we demonstrate an alternative strategy, a growth-coupled, high-yield bioconversion, by feeding cells a non-sugar substrate, by-passing central metabolism, and linking a key metabolic step to generation of acetyl-CoA that is required for biomass and energy generation. Specifically, we converted levulinic acid (LA), an established degradation product of lignocellulosic biomass, to butanone (a.k.a. methyl-ethyl ketone - MEK), a widely used industrial solvent. Our strategy combines a catabolic pathway from Pseudomonas putida that enables conversion of LA to 3-ketovaleryl-CoA, a CoA transferase that generates 3-ketovalerate and acetyl-CoA, and a decarboxylase that generates 2-butanone. By removing the ability of E. coli to consume LA and supplying excess acetate as a carbon source, we built a strain of E. coli that could convert LA to butanone at high yields, but at the cost of significant acetate consumption. Using flux balance analysis as a guide, we built a strain of E. coli that linked acetate assimilation to production of butanone. This strain was capable of complete bioconversion of LA to butanone with a reduced acetate requirement and increased specific productivity. To demonstrate the bioconversion on real world feedstocks, we produced LA from furfuryl alcohol, a compound readily obtained from biomass. These LA feedstocks were found to contain inhibitors that prevented cell growth and bioconversion of LA to butanone. We used a combination of column chromatography and activated carbon to remove the toxic compounds from the feedstock, resulting in LA that could be completely converted to butanone. In conclusion, this work motivates continued collaboration between chemical and biological catalysis researchers to explore alternative conversion pathways and the technical hurdles that prevent their rapid deployment.},
doi = {10.1016/j.ymben.2019.06.003},
journal = {Metabolic Engineering},
number = C,
volume = 55,
place = {United States},
year = {Wed Jun 19 00:00:00 EDT 2019},
month = {Wed Jun 19 00:00:00 EDT 2019}
}
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