Sucrose as an electron source for cofactor regeneration in recombinant Escherichia coli expressing invertase and a Baeyer Villiger monooxygenase.


Journal

Microbial cell factories
ISSN: 1475-2859
Titre abrégé: Microb Cell Fact
Pays: England
ID NLM: 101139812

Informations de publication

Date de publication:
12 Aug 2024
Historique:
received: 08 03 2024
accepted: 08 07 2024
medline: 13 8 2024
pubmed: 13 8 2024
entrez: 12 8 2024
Statut: epublish

Résumé

The large-scale biocatalytic application of oxidoreductases requires systems for a cost-effective and efficient regeneration of redox cofactors. These represent the major bottleneck for industrial bioproduction and an important cost factor. In this work, co-expression of the genes of invertase and a Baeyer-Villiger monooxygenase from Burkholderia xenovorans to E. coli W ΔcscR and E. coli BL21 (DE3) enabled efficient biotransformation of cyclohexanone to the polymer precursor, ε-caprolactone using sucrose as electron source for regeneration of redox cofactors, at rates comparable to glucose. E. coli W ΔcscR has a native csc regulon enabling sucrose utilization and is deregulated via deletion of the repressor gene (cscR), thus enabling sucrose uptake even at concentrations below 6 mM (2 g L Herein, we show a proof of concept where the co-expression of invertase for both E. coli hosts was sufficient for efficient sucrose utilization to sustain cofactor regeneration in the Baeyer-Villiger oxidation of cyclohexanone. Using E. coli W ΔcscR, a specific activity of 37 U g Results show that sucrose can be an alternative electron source to drive whole-cell biotransformations in recombinant E. coli strains opening novel strategies for sustainable chemical production.

Sections du résumé

BACKGROUND BACKGROUND
The large-scale biocatalytic application of oxidoreductases requires systems for a cost-effective and efficient regeneration of redox cofactors. These represent the major bottleneck for industrial bioproduction and an important cost factor. In this work, co-expression of the genes of invertase and a Baeyer-Villiger monooxygenase from Burkholderia xenovorans to E. coli W ΔcscR and E. coli BL21 (DE3) enabled efficient biotransformation of cyclohexanone to the polymer precursor, ε-caprolactone using sucrose as electron source for regeneration of redox cofactors, at rates comparable to glucose. E. coli W ΔcscR has a native csc regulon enabling sucrose utilization and is deregulated via deletion of the repressor gene (cscR), thus enabling sucrose uptake even at concentrations below 6 mM (2 g L
RESULTS RESULTS
Herein, we show a proof of concept where the co-expression of invertase for both E. coli hosts was sufficient for efficient sucrose utilization to sustain cofactor regeneration in the Baeyer-Villiger oxidation of cyclohexanone. Using E. coli W ΔcscR, a specific activity of 37 U g
CONCLUSIONS CONCLUSIONS
Results show that sucrose can be an alternative electron source to drive whole-cell biotransformations in recombinant E. coli strains opening novel strategies for sustainable chemical production.

Identifiants

pubmed: 39135032
doi: 10.1186/s12934-024-02474-2
pii: 10.1186/s12934-024-02474-2
doi:

Substances chimiques

beta-Fructofuranosidase EC 3.2.1.26
Sucrose 57-50-1
Mixed Function Oxygenases EC 1.-
Cyclohexanones 0
cyclohexanone 5QOR3YM052
Recombinant Proteins 0
caprolactone 56RE988L1R
Caproates 0
Lactones 0

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

227

Subventions

Organisme : European Union, FET Open
ID : 899576
Organisme : European Union, FET Open
ID : 899576
Organisme : European Union, FET Open
ID : 899576
Organisme : European Union, FET Open
ID : 899576
Organisme : European Union, FET Open
ID : 899576
Organisme : European Union, FET Open
ID : 899576
Organisme : Novo Nordisk Foundation
ID : NNF20OC0064371

Informations de copyright

© 2024. The Author(s).

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Auteurs

Lucija Sovic (L)

Institute of Molecular Biotechnology, Graz University of Technology, NAWI Graz, Petersgasse 14, 8010, Graz, Austria.

Lenny Malihan-Yap (L)

Institute of Molecular Biotechnology, Graz University of Technology, NAWI Graz, Petersgasse 14, 8010, Graz, Austria.

Gábor Szilveszter Tóth (GS)

Molecular Plant Biology, Department of Life Technologies, University of Turku, 20014, Turku, Finland.

Vilja Siitonen (V)

Molecular Plant Biology, Department of Life Technologies, University of Turku, 20014, Turku, Finland.

Véronique Alphand (V)

Aix Marseille Univ, CNRS, Centrale Marseille, iSm2, Marseille, France.

Yagut Allahverdiyeva (Y)

Molecular Plant Biology, Department of Life Technologies, University of Turku, 20014, Turku, Finland.

Robert Kourist (R)

Institute of Molecular Biotechnology, Graz University of Technology, NAWI Graz, Petersgasse 14, 8010, Graz, Austria. kourist@tugraz.at.
ACIB GmbH, 8010, Graz, Austria. kourist@tugraz.at.

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