Development of an Haa1-based biosensor for acetic acid sensing in Saccharomyces cerevisiae
Journal article, 2021

Acetic acid is one of the main inhibitors of lignocellulosic hydrolysates and acetic acid tolerance is crucial for the development of robust cell factories for conversion of biomass. As a precursor of acetyl-coenzyme A, it also plays an important role in central carbon metabolism. Thus, monitoring acetic acid levels is a crucial aspect when cultivating yeast. Transcription factor-based biosensors represent useful tools to follow metabolite concentrations. Here, we present the development of an acetic acid biosensor based on the Saccharomyces cerevisiae transcription factor Haa1 that upon binding to acetic acid relocates to the nucleus. In the biosensor, a synthetic transcription factor consisting of Haa1 and BM3R1 from Bacillus megaterium was used to control expression of a reporter gene under a promoter containing BM3R1 binding sites. The biosensor did not drive expression under a promoter containing Haa1 binding sites and responded to acetic acid over a linear range spanning from 10 to 60 mM. To validate its applicability, the biosensor was integrated into acetic acid-producing strains. A direct correlation between biosensor output and acetic acid production was detected. The developed biosensor enables high-throughput screening of strains producing acetic acid and could also be used to investigate acetic acid-tolerant strain libraries.

Haa1

biosensor

acetic acid

synthetic transcription factor

Author

Maurizio Mormino

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

FEMS Yeast Research

1567-1356 (ISSN) 1567-1364 (eISSN)

Vol. 21 6 foab049

Subject Categories

Microbiology

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

Other Industrial Biotechnology

DOI

10.1093/femsyr/foab049

PubMed

34477863

More information

Latest update

11/1/2021