FadR-Based Biosensor-Assisted Screening for Genes Enhancing Fatty Acyl-CoA Pools in Saccharomyces cerevisiae

Yasaman Dabirian, Paulo Gonçalves Teixeira, Jens Nielsen, Verena Siewers, Florian David

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Fatty acid-derived compounds have a range of industrial applications, from chemical building blocks to biofuels. Due to the highly dynamic nature of fatty acid metabolism, it is difficult to identify genes modulating fatty acyl-CoA levels using a rational approach. Metabolite biosensors can be used to screen genes from large-scale libraries in vivo in a high throughput manner. Here, a fatty acyl-CoA sensor based on the transcription factor FadR from Escherichia coli was established in Saccharomyces cerevisiae and combined with a gene overexpression library to screen for genes increasing the fatty acyl-CoA pool. Fluorescence-activated cell sorting, followed by data analysis, identified genes enhancing acyl-CoA levels. From these, overexpression of RTC3, GGA2, and LPP1 resulted in about 80% increased fatty alcohol levels. Changes in fatty acid saturation and chain length distribution could also be observed. These results indicate that the use of this acyl-CoA biosensor combined with a gene overexpression library allows for identification of gene targets improving production of fatty acids and derived products.
Original languageEnglish
JournalACS Synthetic Biology
Volume8
Issue number8
Pages (from-to)1788-1800
ISSN2161-5063
DOIs
Publication statusPublished - 2019

Keywords

  • fatty acyl-CoA sensor
  • FadR
  • Saccharomyces cerevisiae
  • fatty acids
  • fatty alcohols

Cite this

Dabirian, Yasaman ; Gonçalves Teixeira, Paulo ; Nielsen, Jens ; Siewers, Verena ; David, Florian. / FadR-Based Biosensor-Assisted Screening for Genes Enhancing Fatty Acyl-CoA Pools in Saccharomyces cerevisiae. In: ACS Synthetic Biology. 2019 ; Vol. 8, No. 8. pp. 1788-1800.
@article{4cd56c5404784ce794898a9b8fa0b865,
title = "FadR-Based Biosensor-Assisted Screening for Genes Enhancing Fatty Acyl-CoA Pools in Saccharomyces cerevisiae",
abstract = "Fatty acid-derived compounds have a range of industrial applications, from chemical building blocks to biofuels. Due to the highly dynamic nature of fatty acid metabolism, it is difficult to identify genes modulating fatty acyl-CoA levels using a rational approach. Metabolite biosensors can be used to screen genes from large-scale libraries in vivo in a high throughput manner. Here, a fatty acyl-CoA sensor based on the transcription factor FadR from Escherichia coli was established in Saccharomyces cerevisiae and combined with a gene overexpression library to screen for genes increasing the fatty acyl-CoA pool. Fluorescence-activated cell sorting, followed by data analysis, identified genes enhancing acyl-CoA levels. From these, overexpression of RTC3, GGA2, and LPP1 resulted in about 80{\%} increased fatty alcohol levels. Changes in fatty acid saturation and chain length distribution could also be observed. These results indicate that the use of this acyl-CoA biosensor combined with a gene overexpression library allows for identification of gene targets improving production of fatty acids and derived products.",
keywords = "fatty acyl-CoA sensor, FadR, Saccharomyces cerevisiae, fatty acids, fatty alcohols",
author = "Yasaman Dabirian and {Gon{\cc}alves Teixeira}, Paulo and Jens Nielsen and Verena Siewers and Florian David",
year = "2019",
doi = "10.1021/acssynbio.9b00118",
language = "English",
volume = "8",
pages = "1788--1800",
journal = "A C S Synthetic Biology",
issn = "2161-5063",
publisher = "American Chemical Society",
number = "8",

}

FadR-Based Biosensor-Assisted Screening for Genes Enhancing Fatty Acyl-CoA Pools in Saccharomyces cerevisiae. / Dabirian, Yasaman; Gonçalves Teixeira, Paulo; Nielsen, Jens; Siewers, Verena; David, Florian.

In: ACS Synthetic Biology, Vol. 8, No. 8, 2019, p. 1788-1800.

Research output: Contribution to journalJournal articleResearchpeer-review

TY - JOUR

T1 - FadR-Based Biosensor-Assisted Screening for Genes Enhancing Fatty Acyl-CoA Pools in Saccharomyces cerevisiae

AU - Dabirian, Yasaman

AU - Gonçalves Teixeira, Paulo

AU - Nielsen, Jens

AU - Siewers, Verena

AU - David, Florian

PY - 2019

Y1 - 2019

N2 - Fatty acid-derived compounds have a range of industrial applications, from chemical building blocks to biofuels. Due to the highly dynamic nature of fatty acid metabolism, it is difficult to identify genes modulating fatty acyl-CoA levels using a rational approach. Metabolite biosensors can be used to screen genes from large-scale libraries in vivo in a high throughput manner. Here, a fatty acyl-CoA sensor based on the transcription factor FadR from Escherichia coli was established in Saccharomyces cerevisiae and combined with a gene overexpression library to screen for genes increasing the fatty acyl-CoA pool. Fluorescence-activated cell sorting, followed by data analysis, identified genes enhancing acyl-CoA levels. From these, overexpression of RTC3, GGA2, and LPP1 resulted in about 80% increased fatty alcohol levels. Changes in fatty acid saturation and chain length distribution could also be observed. These results indicate that the use of this acyl-CoA biosensor combined with a gene overexpression library allows for identification of gene targets improving production of fatty acids and derived products.

AB - Fatty acid-derived compounds have a range of industrial applications, from chemical building blocks to biofuels. Due to the highly dynamic nature of fatty acid metabolism, it is difficult to identify genes modulating fatty acyl-CoA levels using a rational approach. Metabolite biosensors can be used to screen genes from large-scale libraries in vivo in a high throughput manner. Here, a fatty acyl-CoA sensor based on the transcription factor FadR from Escherichia coli was established in Saccharomyces cerevisiae and combined with a gene overexpression library to screen for genes increasing the fatty acyl-CoA pool. Fluorescence-activated cell sorting, followed by data analysis, identified genes enhancing acyl-CoA levels. From these, overexpression of RTC3, GGA2, and LPP1 resulted in about 80% increased fatty alcohol levels. Changes in fatty acid saturation and chain length distribution could also be observed. These results indicate that the use of this acyl-CoA biosensor combined with a gene overexpression library allows for identification of gene targets improving production of fatty acids and derived products.

KW - fatty acyl-CoA sensor

KW - FadR

KW - Saccharomyces cerevisiae

KW - fatty acids

KW - fatty alcohols

U2 - 10.1021/acssynbio.9b00118

DO - 10.1021/acssynbio.9b00118

M3 - Journal article

VL - 8

SP - 1788

EP - 1800

JO - A C S Synthetic Biology

JF - A C S Synthetic Biology

SN - 2161-5063

IS - 8

ER -