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Unraveling the mechanism of furfural tolerance in engineered Pseudomonas putida by genomics

As a dehydration product of pentoses in hemicellulose sugar streams derived from lignocellulosic biomass, furfural is a prevalent inhibitor in the efficient microbial conversion process. To solve this obstacle, exploiting a biorefinery strain with remarkable furfural tolerance capability is essentia...

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Autores principales: Zou, Lihua, Jin, Xinzhu, Tao, Yuanming, Zheng, Zhaojuan, Ouyang, Jia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9630843/
https://www.ncbi.nlm.nih.gov/pubmed/36338095
http://dx.doi.org/10.3389/fmicb.2022.1035263
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author Zou, Lihua
Jin, Xinzhu
Tao, Yuanming
Zheng, Zhaojuan
Ouyang, Jia
author_facet Zou, Lihua
Jin, Xinzhu
Tao, Yuanming
Zheng, Zhaojuan
Ouyang, Jia
author_sort Zou, Lihua
collection PubMed
description As a dehydration product of pentoses in hemicellulose sugar streams derived from lignocellulosic biomass, furfural is a prevalent inhibitor in the efficient microbial conversion process. To solve this obstacle, exploiting a biorefinery strain with remarkable furfural tolerance capability is essential. Pseudomonas putida KT2440 (P. putida) has served as a valuable bacterial chassis for biomass biorefinery. Here, a high-concentration furfural-tolerant P. putida strain was developed via adaptive laboratory evolution (ALE). The ALE resulted in a previously engineered P. putida strain with substantially increased furfural tolerance as compared to wild-type. Whole-genome sequencing of the adapted strains and reverse engineering validation of key targets revealed for the first time that several genes and their mutations, especially for PP_RS19785 and PP_RS18130 [encoding ATP-binding cassette (ABC) transporters] as well as PP_RS20740 (encoding a hypothetical protein), play pivotal roles in the furfural tolerance and conversion of this bacterium. Finally, strains overexpressing these three striking mutations grew well in highly toxic lignocellulosic hydrolysate, with cell biomass around 9-, 3.6-, and two-fold improvement over the control strain, respectively. To our knowledge, this study first unravels the furan aldehydes tolerance mechanism of industrial workhorse P. putida, which provides a new foundation for engineering strains to enhance furfural tolerance and further facilitate the valorization of lignocellulosic biomass.
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spelling pubmed-96308432022-11-04 Unraveling the mechanism of furfural tolerance in engineered Pseudomonas putida by genomics Zou, Lihua Jin, Xinzhu Tao, Yuanming Zheng, Zhaojuan Ouyang, Jia Front Microbiol Microbiology As a dehydration product of pentoses in hemicellulose sugar streams derived from lignocellulosic biomass, furfural is a prevalent inhibitor in the efficient microbial conversion process. To solve this obstacle, exploiting a biorefinery strain with remarkable furfural tolerance capability is essential. Pseudomonas putida KT2440 (P. putida) has served as a valuable bacterial chassis for biomass biorefinery. Here, a high-concentration furfural-tolerant P. putida strain was developed via adaptive laboratory evolution (ALE). The ALE resulted in a previously engineered P. putida strain with substantially increased furfural tolerance as compared to wild-type. Whole-genome sequencing of the adapted strains and reverse engineering validation of key targets revealed for the first time that several genes and their mutations, especially for PP_RS19785 and PP_RS18130 [encoding ATP-binding cassette (ABC) transporters] as well as PP_RS20740 (encoding a hypothetical protein), play pivotal roles in the furfural tolerance and conversion of this bacterium. Finally, strains overexpressing these three striking mutations grew well in highly toxic lignocellulosic hydrolysate, with cell biomass around 9-, 3.6-, and two-fold improvement over the control strain, respectively. To our knowledge, this study first unravels the furan aldehydes tolerance mechanism of industrial workhorse P. putida, which provides a new foundation for engineering strains to enhance furfural tolerance and further facilitate the valorization of lignocellulosic biomass. Frontiers Media S.A. 2022-10-20 /pmc/articles/PMC9630843/ /pubmed/36338095 http://dx.doi.org/10.3389/fmicb.2022.1035263 Text en Copyright © 2022 Zou, Jin, Tao, Zheng and Ouyang. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Zou, Lihua
Jin, Xinzhu
Tao, Yuanming
Zheng, Zhaojuan
Ouyang, Jia
Unraveling the mechanism of furfural tolerance in engineered Pseudomonas putida by genomics
title Unraveling the mechanism of furfural tolerance in engineered Pseudomonas putida by genomics
title_full Unraveling the mechanism of furfural tolerance in engineered Pseudomonas putida by genomics
title_fullStr Unraveling the mechanism of furfural tolerance in engineered Pseudomonas putida by genomics
title_full_unstemmed Unraveling the mechanism of furfural tolerance in engineered Pseudomonas putida by genomics
title_short Unraveling the mechanism of furfural tolerance in engineered Pseudomonas putida by genomics
title_sort unraveling the mechanism of furfural tolerance in engineered pseudomonas putida by genomics
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9630843/
https://www.ncbi.nlm.nih.gov/pubmed/36338095
http://dx.doi.org/10.3389/fmicb.2022.1035263
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