Improved vanillin production in baker's yeast through in silico design

Publication: Research - peer-reviewJournal article – Annual report year: 2010

Standard

Improved vanillin production in baker's yeast through in silico design. / Brochado, Ana Rita; Matos, Cláudia; Møller, Birger L.; Hansen, Jørgen; Mortensen, Uffe H.; Patil, Kiran Raosaheb.

In: Microbial Cell Factories, Vol. 9, 2010, p. 84.

Publication: Research - peer-reviewJournal article – Annual report year: 2010

Harvard

APA

CBE

MLA

Vancouver

Author

Brochado, Ana Rita; Matos, Cláudia; Møller, Birger L.; Hansen, Jørgen; Mortensen, Uffe H.; Patil, Kiran Raosaheb / Improved vanillin production in baker's yeast through in silico design.

In: Microbial Cell Factories, Vol. 9, 2010, p. 84.

Publication: Research - peer-reviewJournal article – Annual report year: 2010

Bibtex

@article{7eeeb11a53674e03b5d6f34785c588c1,
title = "Improved vanillin production in baker's yeast through in silico design",
publisher = "BioMed Central Ltd.",
author = "Brochado, {Ana Rita} and Cláudia Matos and Møller, {Birger L.} and Jørgen Hansen and Mortensen, {Uffe H.} and Patil, {Kiran Raosaheb}",
note = "This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.",
year = "2010",
doi = "10.1186/1475-2859-9-84",
volume = "9",
pages = "84",
journal = "Microbial Cell Factories",
issn = "1475-2859",

}

RIS

TY - JOUR

T1 - Improved vanillin production in baker's yeast through in silico design

A1 - Brochado,Ana Rita

A1 - Matos,Cláudia

A1 - Møller,Birger L.

A1 - Hansen,Jørgen

A1 - Mortensen,Uffe H.

A1 - Patil,Kiran Raosaheb

AU - Brochado,Ana Rita

AU - Matos,Cláudia

AU - Møller,Birger L.

AU - Hansen,Jørgen

AU - Mortensen,Uffe H.

AU - Patil,Kiran Raosaheb

PB - BioMed Central Ltd.

PY - 2010

Y1 - 2010

N2 - Background: Vanillin is one of the most widely used flavouring agents, originally obtained from cured seed pods of the vanilla orchid Vanilla planifolia. Currently vanillin is mostly produced via chemical synthesis. A de novo synthetic pathway for heterologous vanillin production from glucose has recently been implemented in baker's yeast, Saccharamyces cerevisiae. In this study we aimed at engineering this vanillin cell factory towards improved productivity and thereby at developing an attractive alternative to chemical synthesis. Results: Expression of a glycosyltransferase from Arabidopsis thaliana in the vanillin producing S. cerevisiae strain served to decrease product toxicity. An in silico metabolic engineering strategy of this vanillin glucoside producing strain was designed using a set of stoichiometric modelling tools applied to the yeast genome-scale metabolic network. Two targets (PDC1 and GDH1) were selected for experimental verification resulting in four engineered strains. Three of the mutants showed up to 1.5 fold higher vanillin beta-D-glucoside yield in batch mode, while continuous culture of the Delta pdc1 mutant showed a 2-fold productivity improvement. This mutant presented a 5-fold improvement in free vanillin production compared to the previous work on de novo vanillin biosynthesis in baker's yeast. Conclusion: Use of constraints corresponding to different physiological states was found to greatly influence the target predictions given minimization of metabolic adjustment (MOMA) as biological objective function. In vivo verification of the targets, selected based on their predicted metabolic adjustment, successfully led to overproducing strains. Overall, we propose and demonstrate a framework for in silico design and target selection for improving microbial cell factories.

AB - Background: Vanillin is one of the most widely used flavouring agents, originally obtained from cured seed pods of the vanilla orchid Vanilla planifolia. Currently vanillin is mostly produced via chemical synthesis. A de novo synthetic pathway for heterologous vanillin production from glucose has recently been implemented in baker's yeast, Saccharamyces cerevisiae. In this study we aimed at engineering this vanillin cell factory towards improved productivity and thereby at developing an attractive alternative to chemical synthesis. Results: Expression of a glycosyltransferase from Arabidopsis thaliana in the vanillin producing S. cerevisiae strain served to decrease product toxicity. An in silico metabolic engineering strategy of this vanillin glucoside producing strain was designed using a set of stoichiometric modelling tools applied to the yeast genome-scale metabolic network. Two targets (PDC1 and GDH1) were selected for experimental verification resulting in four engineered strains. Three of the mutants showed up to 1.5 fold higher vanillin beta-D-glucoside yield in batch mode, while continuous culture of the Delta pdc1 mutant showed a 2-fold productivity improvement. This mutant presented a 5-fold improvement in free vanillin production compared to the previous work on de novo vanillin biosynthesis in baker's yeast. Conclusion: Use of constraints corresponding to different physiological states was found to greatly influence the target predictions given minimization of metabolic adjustment (MOMA) as biological objective function. In vivo verification of the targets, selected based on their predicted metabolic adjustment, successfully led to overproducing strains. Overall, we propose and demonstrate a framework for in silico design and target selection for improving microbial cell factories.

U2 - 10.1186/1475-2859-9-84

DO - 10.1186/1475-2859-9-84

JO - Microbial Cell Factories

JF - Microbial Cell Factories

SN - 1475-2859

VL - 9

SP - 84

ER -