Identification of acetic acid sensitive strains through biosensor-based screening of a Saccharomyces cerevisiae CRISPRi library
Journal article, 2022

BACKGROUND: Acetic acid tolerance is crucial for the development of robust cell factories for conversion of lignocellulosic hydrolysates that typically contain high levels of acetic acid. Screening mutants for growth in medium with acetic acid is an attractive way to identify sensitive variants and can provide novel insights into the complex mechanisms regulating the acetic acid stress response. RESULTS: An acetic acid biosensor based on the Saccharomyces cerevisiae transcription factor Haa1, was used to screen a CRISPRi yeast strain library where dCas9-Mxi was set to individually repress each essential or respiratory growth essential gene. Fluorescence-activated cell sorting led to the enrichment of a population of cells with higher acetic acid retention. These cells with higher biosensor signal were demonstrated to be more sensitive to acetic acid. Biosensor-based screening of the CRISPRi library strains enabled identification of strains with increased acetic acid sensitivity: strains with gRNAs targeting TIF34, MSN5, PAP1, COX10 or TRA1. CONCLUSIONS: This study demonstrated that biosensors are valuable tools for screening and monitoring acetic acid tolerance in yeast. Fine-tuning the expression of essential genes can lead to altered acetic acid tolerance.

Yeast

Tolerance

Screening

CRISPRi

Library

Acetic acid

Biosensor

Author

Maurizio Mormino

Chalmers, Biology and Biological Engineering, Industrial Biotechnology

Ibai Lenitz Etxaburu

Chalmers, Biology and Biological Engineering, Industrial Biotechnology

Verena Siewers

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Yvonne Nygård

Chalmers, Biology and Biological Engineering, Industrial Biotechnology

Microbial Cell Factories

14752859 (eISSN)

Vol. 21 1 214-

Genetic biosensors for developing better cell factories for production of platform chemicals

Adlerbertska Foundations, 2018-01-01 -- 2021-12-31.

ÅForsk, 2018-01-01 -- 2021-12-31.

Ollie och Elof Ericssons stiftelser, 2018-01-01 -- 2022-10-31.

Formas (2017-00979), 2018-01-01 -- 2020-12-31.

Subject Categories

Cell Biology

Cell and Molecular Biology

Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)

DOI

10.1186/s12934-022-01938-7

PubMed

36243715

Related datasets

Additional file 2 of Identification of acetic acid sensitive strains through biosensor-based screening of a Saccharomyces cerevisiae CRISPRi library [dataset]

DOI: 10.6084/m9.figshare.21343096

More information

Latest update

5/4/2023 7