title: Development of a Synthetic Malonyl-CoA Sensor in Saccharomyces cerevisiae for Intracellular Metabolite Monitoring and Genetic Screening.
abstract: Genetic sensors capable of converting key metabolite levels to fluorescence signals enable the monitoring of intracellular compound concentrations in living cells, and emerge as an efficient tool in high-throughput genetic screening. However, the development of genetic sensors in yeasts lags far behind their development in bacteria. Here we report the design of a malonyl-CoA sensor in Saccharomyces cerevisiae using an adapted bacterial transcription factor FapR and its corresponding operator fapO to gauge intracellular malonyl-CoA levels. By combining this sensor with a genome-wide overexpression library, we identified two novel gene targets that improved intracellular malonyl-CoA concentration. We further utilized the resulting recombinant yeast strain to produce a valuable compound, 3-hydroxypropionic acid, from malonyl-CoA and enhanced its titer by 120%. Such a genetic sensor provides a powerful approach for genome-wide screening and could further improve the synthesis of a large range of chemicals derived from malonyl-CoA in yeast.
authors.affiliation: Energy Biosciences Institute, ‡Institute for Genomic Biology, §Department of Chemical and Biomolecular Engineering, and ∥Departments of Chemistry, Biochemistry, and Bioengineering, University of Illinois at Urbana-Champaign , Urbana, Illinois 61801, United States.
authors.forename: Sijin
authors.initials: S
authors.lastname: Li
authors.affiliation: Energy Biosciences Institute, ‡Institute for Genomic Biology, §Department of Chemical and Biomolecular Engineering, and ∥Departments of Chemistry, Biochemistry, and Bioengineering, University of Illinois at Urbana-Champaign , Urbana, Illinois 61801, United States.
authors.forename: Tong
authors.initials: T
authors.lastname: Si
authors.affiliation: Energy Biosciences Institute, ‡Institute for Genomic Biology, §Department of Chemical and Biomolecular Engineering, and ∥Departments of Chemistry, Biochemistry, and Bioengineering, University of Illinois at Urbana-Champaign , Urbana, Illinois 61801, United States.
authors.forename: Meng
authors.initials: M
authors.lastname: Wang
authors.affiliation: Energy Biosciences Institute, ‡Institute for Genomic Biology, §Department of Chemical and Biomolecular Engineering, and ∥Departments of Chemistry, Biochemistry, and Bioengineering, University of Illinois at Urbana-Champaign , Urbana, Illinois 61801, United States.
authors.forename: Huimin
authors.initials: H
authors.lastname: Zhao
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D029968:Escherichia coli Proteins / Q000096:biosynthesis / Q000235:genetics; D008316:Malonyl Coenzyme A / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D009875:Operator Regions, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000096:biosynthesis / Q000235:genetics
chemical_list: D029968:Escherichia coli Proteins; C067955:FAPR protein, E coli; D014157:Transcription Factors; D008316:Malonyl Coenzyme A
keywords: 3-hydroxypropionic acid; genetic sensor; genome-wide library; high-throughput screening; malonyl-CoA