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Rational and evolut...
Rational and evolutionary engineering of Saccharomyces cerevisiae for production of dicarboxylic acids from lignocellulosic biomass and exploring genetic mechanisms of the yeast tolerance to the biomass hydrolysate
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- Stovicek, Vratislav (författare)
- Danmarks Tekniske Universitet,Technical University of Denmark
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- Dato, Laura (författare)
- Danmarks Tekniske Universitet,Technical University of Denmark
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- Almqvist, Henrik (författare)
- Lund University,Lunds universitet,Avdelningen för kemiteknik,Institutionen för processteknik och tillämpad biovetenskap,Institutioner vid LTH,Lunds Tekniska Högskola,Division of Chemical Engineering,Department of Process and Life Science Engineering,Departments at LTH,Faculty of Engineering, LTH
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visa fler...
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- Schöpping, Marie, 1991 (författare)
- Chalmers University of Technology,Lund University,Lunds universitet,Avdelningen för kemiteknik,Institutionen för processteknik och tillämpad biovetenskap,Institutioner vid LTH,Lunds Tekniska Högskola,Division of Chemical Engineering,Department of Process and Life Science Engineering,Departments at LTH,Faculty of Engineering, LTH,Chr. Hansen Holding A/S
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- Chekina, Ksenia (författare)
- Danmarks Tekniske Universitet,Technical University of Denmark
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- Pedersen, Lasse Ebdrup (författare)
- Danmarks Tekniske Universitet,Technical University of Denmark
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- Koza, Anna (författare)
- Chr. Hansen Holding A/S,Danmarks Tekniske Universitet,Technical University of Denmark
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- Figueira, Diogo (författare)
- Biotrend S.A.,Biotrend
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- Tjosås, Freddy (författare)
- Borregaard AS
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- Ferreira, Bruno Sommer (författare)
- Biotrend S.A.,Biotrend
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- Forster, J. (författare)
- Danmarks Tekniske Universitet,Technical University of Denmark
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- Lidén, Gunnar (författare)
- Lund University,Lunds universitet,Avdelningen för kemiteknik,Institutionen för processteknik och tillämpad biovetenskap,Institutioner vid LTH,Lunds Tekniska Högskola,Division of Chemical Engineering,Department of Process and Life Science Engineering,Departments at LTH,Faculty of Engineering, LTH
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- Borodina, I. (författare)
- Danmarks Tekniske Universitet,Technical University of Denmark
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(creator_code:org_t)
- 2022-02-27
- 2022
- Engelska.
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Ingår i: Biotechnology for Biofuels and Bioproducts. - : Springer Science and Business Media LLC. - 2731-3654. ; 15:1
- Relaterad länk:
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https://research.cha... (primary) (free)
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https://biotechnolog...
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http://dx.doi.org/10... (free)
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https://research.cha...
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https://doi.org/10.1...
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https://lup.lub.lu.s...
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Abstract
Ämnesord
Stäng
- Background: Lignosulfonates are significant wood chemicals with a $700 million market, produced by sulfite pulping of wood. During the pulping process, spent sulfite liquor (SSL) is generated, which in addition to lignosulfonates contains hemicellulose-derived sugars—in case of hardwoods primarily the pentose sugar xylose. The pentoses are currently underutilized. If they could be converted into value-added chemicals, overall economic profitability of the process would increase. SSLs are typically very inhibitory to microorganisms, which presents a challenge for a biotechnological process. The aim of the present work was to develop a robust yeast strain able to convert xylose in SSL to carboxylic acids. Results: The industrial strain Ethanol Red of the yeast Saccharomyces cerevisiae was engineered for efficient utilization of xylose in a Eucalyptus globulus lignosulfonate stream at low pH using CRISPR/Cas genome editing and adaptive laboratory evolution. The engineered strain grew in synthetic medium with xylose as sole carbon source with maximum specific growth rate (µmax) of 0.28 1/h. Selected evolved strains utilized all carbon sources in the SSL at pH 3.5 and grew with µmax between 0.05 and 0.1 1/h depending on a nitrogen source supplement. Putative genetic determinants of the increased tolerance to the SSL were revealed by whole genome sequencing of the evolved strains. In particular, four top-candidate genes (SNG1, FIT3, FZF1 and CBP3) were identified along with other gene candidates with predicted important roles, based on the type and distribution of the mutations across different strains and especially the best performing ones. The developed strains were further engineered for production of dicarboxylic acids (succinic and malic acid) via overexpression of the reductive branch of the tricarboxylic acid cycle (TCA). The production strain produced 0.2 mol and 0.12 mol of malic acid and succinic acid, respectively, per mol of xylose present in the SSL. Conclusions: The combined metabolic engineering and adaptive evolution approach provided a robust SSL-tolerant industrial strain that converts fermentable carbon content of the SSL feedstock into malic and succinic acids at low pH.in production yields reaching 0.1 mol and 0.065 mol per mol of total consumed carbon sources. Moreover, our work suggests potential genetic background of the tolerance to the SSL stream pointing out potential gene targets for improving the tolerance to inhibitory industrial feedstocks.
Ämnesord
- TEKNIK OCH TEKNOLOGIER -- Kemiteknik -- Kemiska processer (hsv//swe)
- ENGINEERING AND TECHNOLOGY -- Chemical Engineering -- Chemical Process Engineering (hsv//eng)
- NATURVETENSKAP -- Biologi -- Mikrobiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Microbiology (hsv//eng)
- TEKNIK OCH TEKNOLOGIER -- Industriell bioteknik -- Biokatalys och enzymteknik (hsv//swe)
- ENGINEERING AND TECHNOLOGY -- Industrial Biotechnology -- Biocatalysis and Enzyme Technology (hsv//eng)
- TEKNIK OCH TEKNOLOGIER -- Industriell bioteknik -- Bioenergi (hsv//swe)
- ENGINEERING AND TECHNOLOGY -- Industrial Biotechnology -- Bioenergy (hsv//eng)
Nyckelord
- Dicarboxylic acids
- Next generation sequencing
- Hardwood spent sulfite liquor
- Xylose
- Industrial yeast
- Biorefineries
- Adaptive evolution
- Adaptive evolution
- Biorefineries
- Dicarboxylic acids
- Hardwood spent sulfite liquor
- Industrial yeast
- Next generation sequencing
- Xylose
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Stovicek, Vratis ...
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Dato, Laura
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Almqvist, Henrik
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Schöpping, Marie ...
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Chekina, Ksenia
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Pedersen, Lasse ...
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Koza, Anna
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Figueira, Diogo
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Tjosås, Freddy
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Ferreira, Bruno ...
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Forster, J.
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Lidén, Gunnar
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Borodina, I.
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- TEKNIK OCH TEKNOLOGIER
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TEKNIK OCH TEKNO ...
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och Kemiteknik
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- NATURVETENSKAP
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NATURVETENSKAP
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och Mikrobiologi
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TEKNIK OCH TEKNO ...
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och Biokatalys och e ...
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