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Metabolic engineering of Clostridium autoethanogenum for ethyl acetate production from CO

BACKGROUND: Ethyl acetate is a bulk chemical traditionally produced via energy intensive chemical esterification. Microbial production of this compound offers promise as a more sustainable alternative process. So far, efforts have focused on using sugar-based feedstocks for microbial ester productio...

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Autores principales: Dykstra, James C., van Oort, Jelle, Yazdi, Ali Tafazoli, Vossen, Eric, Patinios, Constantinos, van der Oost, John, Sousa, Diana Z., Kengen, Servé W. M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9686113/
https://www.ncbi.nlm.nih.gov/pubmed/36419165
http://dx.doi.org/10.1186/s12934-022-01964-5
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author Dykstra, James C.
van Oort, Jelle
Yazdi, Ali Tafazoli
Vossen, Eric
Patinios, Constantinos
van der Oost, John
Sousa, Diana Z.
Kengen, Servé W. M.
author_facet Dykstra, James C.
van Oort, Jelle
Yazdi, Ali Tafazoli
Vossen, Eric
Patinios, Constantinos
van der Oost, John
Sousa, Diana Z.
Kengen, Servé W. M.
author_sort Dykstra, James C.
collection PubMed
description BACKGROUND: Ethyl acetate is a bulk chemical traditionally produced via energy intensive chemical esterification. Microbial production of this compound offers promise as a more sustainable alternative process. So far, efforts have focused on using sugar-based feedstocks for microbial ester production, but extension to one-carbon substrates, such as CO and CO(2)/H(2), is desirable. Acetogens present a promising microbial platform for the production of ethyl esters from these one-carbon substrates. RESULTS: We engineered the acetogen C. autoethanogenum to produce ethyl acetate from CO by heterologous expression of an alcohol acetyltransferase (AAT), which catalyzes the formation of ethyl acetate from acetyl-CoA and ethanol. Two AATs, Eat1 from Kluyveromyces marxianus and Atf1 from Saccharomyces cerevisiae, were expressed in C. autoethanogenum. Strains expressing Atf1 produced up to 0.2 mM ethyl acetate. Ethyl acetate production was barely detectable (< 0.01 mM) for strains expressing Eat1. Supplementation of ethanol was investigated as potential boost for ethyl acetate production but resulted only in a 1.5-fold increase (0.3 mM ethyl acetate). Besides ethyl acetate, C. autoethanogenum expressing Atf1 could produce 4.5 mM of butyl acetate when 20 mM butanol was supplemented to the growth medium. CONCLUSIONS: This work offers for the first time a proof-of-principle that autotrophic short chain ester production from C1-carbon feedstocks is possible and offers leads on how this approach can be optimized in the future. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12934-022-01964-5.
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spelling pubmed-96861132022-11-25 Metabolic engineering of Clostridium autoethanogenum for ethyl acetate production from CO Dykstra, James C. van Oort, Jelle Yazdi, Ali Tafazoli Vossen, Eric Patinios, Constantinos van der Oost, John Sousa, Diana Z. Kengen, Servé W. M. Microb Cell Fact Research BACKGROUND: Ethyl acetate is a bulk chemical traditionally produced via energy intensive chemical esterification. Microbial production of this compound offers promise as a more sustainable alternative process. So far, efforts have focused on using sugar-based feedstocks for microbial ester production, but extension to one-carbon substrates, such as CO and CO(2)/H(2), is desirable. Acetogens present a promising microbial platform for the production of ethyl esters from these one-carbon substrates. RESULTS: We engineered the acetogen C. autoethanogenum to produce ethyl acetate from CO by heterologous expression of an alcohol acetyltransferase (AAT), which catalyzes the formation of ethyl acetate from acetyl-CoA and ethanol. Two AATs, Eat1 from Kluyveromyces marxianus and Atf1 from Saccharomyces cerevisiae, were expressed in C. autoethanogenum. Strains expressing Atf1 produced up to 0.2 mM ethyl acetate. Ethyl acetate production was barely detectable (< 0.01 mM) for strains expressing Eat1. Supplementation of ethanol was investigated as potential boost for ethyl acetate production but resulted only in a 1.5-fold increase (0.3 mM ethyl acetate). Besides ethyl acetate, C. autoethanogenum expressing Atf1 could produce 4.5 mM of butyl acetate when 20 mM butanol was supplemented to the growth medium. CONCLUSIONS: This work offers for the first time a proof-of-principle that autotrophic short chain ester production from C1-carbon feedstocks is possible and offers leads on how this approach can be optimized in the future. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12934-022-01964-5. BioMed Central 2022-11-23 /pmc/articles/PMC9686113/ /pubmed/36419165 http://dx.doi.org/10.1186/s12934-022-01964-5 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Dykstra, James C.
van Oort, Jelle
Yazdi, Ali Tafazoli
Vossen, Eric
Patinios, Constantinos
van der Oost, John
Sousa, Diana Z.
Kengen, Servé W. M.
Metabolic engineering of Clostridium autoethanogenum for ethyl acetate production from CO
title Metabolic engineering of Clostridium autoethanogenum for ethyl acetate production from CO
title_full Metabolic engineering of Clostridium autoethanogenum for ethyl acetate production from CO
title_fullStr Metabolic engineering of Clostridium autoethanogenum for ethyl acetate production from CO
title_full_unstemmed Metabolic engineering of Clostridium autoethanogenum for ethyl acetate production from CO
title_short Metabolic engineering of Clostridium autoethanogenum for ethyl acetate production from CO
title_sort metabolic engineering of clostridium autoethanogenum for ethyl acetate production from co
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9686113/
https://www.ncbi.nlm.nih.gov/pubmed/36419165
http://dx.doi.org/10.1186/s12934-022-01964-5
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