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.

Adaptive evolution Biorefineries Dicarboxylic acids Hardwood spent sulfite liquor Industrial yeast Next generation sequencing Xylose

Journal

Biotechnology for biofuels and bioproducts
ISSN: 2731-3654
Titre abrégé: Biotechnol Biofuels Bioprod
Pays: England
ID NLM: 9918300888906676

Informations de publication

Date de publication:
27 Feb 2022
Historique:
received: 11 11 2021
accepted: 12 02 2022
entrez: 27 2 2022
pubmed: 28 2 2022
medline: 28 2 2022
Statut: epublish

Résumé

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. 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 (µ 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.

Sections du résumé

BACKGROUND 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 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 (µ
CONCLUSIONS 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.

Identifiants

pubmed: 35219341
doi: 10.1186/s13068-022-02121-1
pii: 10.1186/s13068-022-02121-1
pmc: PMC8882276
doi:

Types de publication

Journal Article

Langues

eng

Pagination

22

Subventions

Organisme : Seventh Framework Programme
ID : FP7-613771
Organisme : Novo Nordisk Fonden
ID : NNF20OC0060809
Organisme : Novo Nordisk Fonden
ID : NNF20CC0035580
Organisme : H2020 European Research Council
ID : 757384

Informations de copyright

© 2022. The Author(s).

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Auteurs

Vratislav Stovicek (V)

The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kemitorvet, Building 220, 2800, Kgs. Lyngby, Denmark.

Laura Dato (L)

The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kemitorvet, Building 220, 2800, Kgs. Lyngby, Denmark.
River Stone Biotech ApS, Fruebjergvej 3, 2100, Copenhagen, Denmark.

Henrik Almqvist (H)

Department of Chemical Engineering, Lund University, P.O. Box 124, 221 00, Lund, Sweden.

Marie Schöpping (M)

Department of Chemical Engineering, Lund University, P.O. Box 124, 221 00, Lund, Sweden.
Chr. Hansen A/S, Boge Alle 10-12, 2970, Hørsholm, Denmark.
Division of Industrial Biotechnology, Department of Biology and Biological Engineering, Chalmers University of Technology, 412 96, Gothenburg, Sweden.

Ksenia Chekina (K)

The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kemitorvet, Building 220, 2800, Kgs. Lyngby, Denmark.

Lasse Ebdrup Pedersen (LE)

The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kemitorvet, Building 220, 2800, Kgs. Lyngby, Denmark.

Anna Koza (A)

The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kemitorvet, Building 220, 2800, Kgs. Lyngby, Denmark.
Chr. Hansen A/S, Boge Alle 10-12, 2970, Hørsholm, Denmark.

Diogo Figueira (D)

Biotrend S.A., Biocant Park Núcleo 04, Lote 2, 3060-197, Cantanhede, Portugal.

Freddy Tjosås (F)

Borregaard ApS, Hjalmar Wessels vei 6, 1721, Sarpsborg, Norway.

Bruno Sommer Ferreira (BS)

Biotrend S.A., Biocant Park Núcleo 04, Lote 2, 3060-197, Cantanhede, Portugal.

Jochen Forster (J)

The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kemitorvet, Building 220, 2800, Kgs. Lyngby, Denmark.

Gunnar Lidén (G)

Department of Chemical Engineering, Lund University, P.O. Box 124, 221 00, Lund, Sweden.

Irina Borodina (I)

The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kemitorvet, Building 220, 2800, Kgs. Lyngby, Denmark. irbo@biosustain.dtu.dk.

Classifications MeSH