Efficient sex hormone biosensors in Saccharomyces cerevisiae cells to evaluate human aromatase activity and inhibition
Abstract Yeast sex-hormone whole-cell biosensors are analytical tools characterized by long-time storage and low production cost. We engineered compact β-estradiol biosensors in S. cerevisiae cells by leveraging short (20-nt long) operators bound by the fusion protein LexA-ER-VP64—where ER is the hu...
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Nature Portfolio
2025-01-01
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| Series: | Scientific Reports |
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| Online Access: | https://doi.org/10.1038/s41598-024-85022-7 |
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| author | Jie Wu Matthias Bureik Mario Andrea Marchisio |
| author_facet | Jie Wu Matthias Bureik Mario Andrea Marchisio |
| author_sort | Jie Wu |
| collection | DOAJ |
| description | Abstract Yeast sex-hormone whole-cell biosensors are analytical tools characterized by long-time storage and low production cost. We engineered compact β-estradiol biosensors in S. cerevisiae cells by leveraging short (20-nt long) operators bound by the fusion protein LexA-ER-VP64—where ER is the human estrogen receptor and VP64 a strong viral activation domain. Our best biosensors showed high accuracy since their recovery concentration ranged between 97.13% and 104.69%. As a novelty, we built on top of them testosterone biosensors that exploit the conversion of testosterone into β-estradiol by the human aromatase enzyme—expressed in S. cerevisiae together with its co-factor CPR. We used our engineered yeast strains to evaluate aromatase activity through fluorescence measurements without the need for protein purification. Besides, we set up an aromatase-inhibitors evaluation assay to measure the IC50 (half-maximal inhibitory concentration) of candidate inhibitory compounds and developed a screening assay for enzymes that metabolize β-estradiol that demands only to measure fluorescence. These two assays allow the screening of a large number of chemicals and proteins in a fast and economic fashion. We think that our work will facilitate considerably high throughput screening for the discovery of new drugs and unknown metabolic processes. |
| format | Article |
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| institution | Kabale University |
| issn | 2045-2322 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
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| spelling | doaj-art-febdf49a7305495f94944c41e5451d072025-01-05T12:19:35ZengNature PortfolioScientific Reports2045-23222025-01-0115111510.1038/s41598-024-85022-7Efficient sex hormone biosensors in Saccharomyces cerevisiae cells to evaluate human aromatase activity and inhibitionJie Wu0Matthias Bureik1Mario Andrea Marchisio2School of Pharmaceutical Science and Technology, Tianjin UniversitySchool of Pharmaceutical Science and Technology, Tianjin UniversitySchool of Pharmaceutical Science and Technology, Tianjin UniversityAbstract Yeast sex-hormone whole-cell biosensors are analytical tools characterized by long-time storage and low production cost. We engineered compact β-estradiol biosensors in S. cerevisiae cells by leveraging short (20-nt long) operators bound by the fusion protein LexA-ER-VP64—where ER is the human estrogen receptor and VP64 a strong viral activation domain. Our best biosensors showed high accuracy since their recovery concentration ranged between 97.13% and 104.69%. As a novelty, we built on top of them testosterone biosensors that exploit the conversion of testosterone into β-estradiol by the human aromatase enzyme—expressed in S. cerevisiae together with its co-factor CPR. We used our engineered yeast strains to evaluate aromatase activity through fluorescence measurements without the need for protein purification. Besides, we set up an aromatase-inhibitors evaluation assay to measure the IC50 (half-maximal inhibitory concentration) of candidate inhibitory compounds and developed a screening assay for enzymes that metabolize β-estradiol that demands only to measure fluorescence. These two assays allow the screening of a large number of chemicals and proteins in a fast and economic fashion. We think that our work will facilitate considerably high throughput screening for the discovery of new drugs and unknown metabolic processes.https://doi.org/10.1038/s41598-024-85022-7BiosensorCYP enzymeβ-estradiolTestosteroneAromatase |
| spellingShingle | Jie Wu Matthias Bureik Mario Andrea Marchisio Efficient sex hormone biosensors in Saccharomyces cerevisiae cells to evaluate human aromatase activity and inhibition Scientific Reports Biosensor CYP enzyme β-estradiol Testosterone Aromatase |
| title | Efficient sex hormone biosensors in Saccharomyces cerevisiae cells to evaluate human aromatase activity and inhibition |
| title_full | Efficient sex hormone biosensors in Saccharomyces cerevisiae cells to evaluate human aromatase activity and inhibition |
| title_fullStr | Efficient sex hormone biosensors in Saccharomyces cerevisiae cells to evaluate human aromatase activity and inhibition |
| title_full_unstemmed | Efficient sex hormone biosensors in Saccharomyces cerevisiae cells to evaluate human aromatase activity and inhibition |
| title_short | Efficient sex hormone biosensors in Saccharomyces cerevisiae cells to evaluate human aromatase activity and inhibition |
| title_sort | efficient sex hormone biosensors in saccharomyces cerevisiae cells to evaluate human aromatase activity and inhibition |
| topic | Biosensor CYP enzyme β-estradiol Testosterone Aromatase |
| url | https://doi.org/10.1038/s41598-024-85022-7 |
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