Enhancing n-Butanol Tolerance of Escherichia coli by Overexpressing of Stress-Responsive Molecular Chaperones

Xu, Guochao; Xiao, Lin; Wu, Anning; Han, Ruizhi; Ni, Ye

doi:10.1007/s12010-020-03417-4

Cited by 7 publications

(3 citation statements)

References 41 publications

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“…Therefore, overexpression of Hfq is another interesting strategy that would be worth testing. Recently, overexpression of several sRNA regulators was used to improve butanol-tolerance of E. coli ( Xu et al, 2020 ). A further obvious strategy concerns overexpression of proteins involved in maintaining homeostasis of the transmembrane Δψ and of ATP concentration such as the V-type (Clocel_1654-1662) and F-type (Clocel_3048-3055) ATPases identified in this study.…”

Section: Discussionmentioning

confidence: 99%

“…The development of strains with superior solvent tolerance is highly desirable for sustainable production of biofuels ( Nicolaou et al, 2010 ). Enhancement of microbial solvent tolerance can be achieved through different strategies belonging to two main paradigms: (i) “random” approaches, such as random mutagenesis ( Liu et al, 2012 ), whole genome shuffling ( Mao et al, 2010 ) and adaptive laboratory evolution ( Liu et al, 2013b ) and; (ii) “rational” approaches based on targeted gene modification (e.g., overexpression of genes involved in solvent tolerance) ( Tomas et al, 2003 ; Borden and Papoutsakis, 2007 ; Xu et al, 2020 ).…”

Section: Introductionmentioning

confidence: 99%

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Clostridium cellulovorans Proteomic Responses to Butanol Stress

Costa

Usai

et al. 2021

Front. Microbiol.

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Combination of butanol-hyperproducing and hypertolerant phenotypes is essential for developing microbial strains suitable for industrial production of bio-butanol, one of the most promising liquid biofuels. Clostridium cellulovorans is among the microbial strains with the highest potential for direct production of n-butanol from lignocellulosic wastes, a process that would significantly reduce the cost of bio-butanol. However, butanol exhibits higher toxicity compared to ethanol and C. cellulovorans tolerance to this solvent is low. In the present investigation, comparative gel-free proteomics was used to study the response of C. cellulovorans to butanol challenge and understand the tolerance mechanisms activated in this condition. Sequential Window Acquisition of all Theoretical fragment ion spectra Mass Spectrometry (SWATH-MS) analysis allowed identification and quantification of differentially expressed soluble proteins. The study data are available via ProteomeXchange with the identifier PXD024183. The most important response concerned modulation of protein biosynthesis, folding and degradation. Coherent with previous studies on other bacteria, several heat shock proteins (HSPs), involved in protein quality control, were up-regulated such as the chaperones GroES (Cpn10), Hsp90, and DnaJ. Globally, our data indicate that protein biosynthesis is reduced, likely not to overload HSPs. Several additional metabolic adaptations were triggered by butanol exposure such as the up-regulation of V- and F-type ATPases (involved in ATP synthesis/generation of proton motive force), enzymes involved in amino acid (e.g., arginine, lysine, methionine, and branched chain amino acids) biosynthesis and proteins involved in cell envelope re-arrangement (e.g., the products of Clocel_4136, Clocel_4137, Clocel_4144, Clocel_4162 and Clocel_4352, involved in the biosynthesis of saturated fatty acids) and a redistribution of carbon flux through fermentative pathways (acetate and formate yields were increased and decreased, respectively). Based on these experimental findings, several potential gene targets for metabolic engineering strategies aimed at improving butanol tolerance in C. cellulovorans are suggested. This includes overexpression of HSPs (e.g., GroES, Hsp90, DnaJ, ClpC), RNA chaperone Hfq, V- and F-type ATPases and a number of genes whose function in C. cellulovorans is currently unknown.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Clostridium cellulovorans Proteomic Responses to Butanol Stress

Costa

Usai

et al. 2021

Front. Microbiol.

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“…The availability of genome‐wide engineering techniques such as multiplex automated genome engineering could suit these complex gene modifications (Si et al, 2017 ). Alternatively, strategies targeting global gene regulators involved in stress response could potentially better exploit the native regulatory networks evolved by microorganisms to face these growth conditions (Jones et al, 2016 ; Xu et al, 2021 ). For instance, a number of recent reports indicated an important role of small non‐coding RNAs (sRNAs) and RNA chaperones (e.g., Hfq) in tolerance to a variety of stresses, including butanol, in different microorganisms (Costa et al, 2021 ; Jones et al, 2016 ; Sun et al, 2017 ; Venkataramanan et al, 2013 ).…”

Section: Strategies For Improving Microbial Tolerance To Butanolmentioning

confidence: 99%

Current progress on engineering microbial strains and consortia for production of cellulosic butanol through consolidated bioprocessing

Mazzoli

2022

Microbial Biotechnology

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In the last decades, fermentative production of n‐butanol has regained substantial interest mainly owing to its use as drop‐in‐fuel. The use of lignocellulose as an alternative to traditional acetone–butanol–ethanol fermentation feedstocks (starchy biomass and molasses) can significantly increase the economic competitiveness of biobutanol over production from non‐renewable sources (petroleum). However, the low cost of lignocellulose is offset by its high recalcitrance to biodegradation which generally requires chemical‐physical pre‐treatment and multiple bioreactor‐based processes. The development of consolidated processing (i.e., single‐pot fermentation) can dramatically reduce lignocellulose fermentation costs and promote its industrial application. Here, strategies for developing microbial strains and consortia that feature both efficient (hemi)cellulose depolymerization and butanol production will be depicted, that is, rational metabolic engineering of native (hemi)cellulolytic or native butanol‐producing or other suitable microorganisms; protoplast fusion of (hemi)cellulolytic and butanol‐producing strains; and co‐culture of (hemi)cellulolytic and butanol‐producing microbes. Irrespective of the fermentation feedstock, biobutanol production is inherently limited by the severe toxicity of this solvent that challenges process economic viability. Hence, an overview of strategies for developing butanol hypertolerant strains will be provided.

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