Development of a Synthetic Malonyl-CoA Sensor in Saccharomyces cerevisiae for Intracellular Metabolite Monitoring and Genetic Screening

• Published in: ACS synthetic biology 2015-12, Vol.4 (12), p.1308-1315
• Main Authors: Li, Sijin, Si, Tong, Wang, Meng, Zhao, Huimin
• Format: Article
• Language: English
• Subjects: Biosensing Techniques - methods; Escherichia coli Proteins - biosynthesis; Escherichia coli Proteins - genetics; Malonyl Coenzyme A - analysis; Malonyl Coenzyme A - genetics; Malonyl Coenzyme A - metabolism; Operator Regions, Genetic; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae - metabolism; Transcription Factors - biosynthesis; Transcription Factors - genetics
• ISSN: 2161-5063; 2161-5063
• DOI: 10.1021/acssynbio.5b00069

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.