Journal: Scientific Reports

Article Title: Engineering a GPCR-based yeast biosensor for a highly sensitive melatonin detection from fermented beverages

doi: 10.1038/s41598-024-68633-y

Figure Lengend Snippet: Original biosensor features. ( A ) GPCR integration cassette, this strain has MTNR1A integrated in LEU2 locus. ( B ) Reporter integration cassette bearing a synthetic transcription factor (LexA-PRD), a synthetic promoter driving the expression of the reporter gene sfGFP and a Gα (GPA1) overexpression. This cassette is integrated in URA3 locus. ( C ) Curve fitting of yWS1544 strain derived from Shaw et al. . Measurements are normalized sfGFP fluorescence levels and shown as mean ± SD of triplicates. Curve fitting and parameters were obtained using GraphPad Prism 9.5.1 variable slope (four parameter) nonlinear regression fit.

Article Snippet: Yeast biosensor strain yRB1012 was employed in this assay. sfGFP fluorescence for each sample was interpolated in the fitted calibration curve for melatonin using GraphPad Prism 9.5.1 standard curve interpolation (four parameter) tool.

Techniques: Expressing, Over Expression, Derivative Assay, Fluorescence

Journal: Scientific Reports

Article Title: Engineering a GPCR-based yeast biosensor for a highly sensitive melatonin detection from fermented beverages

doi: 10.1038/s41598-024-68633-y

Figure Lengend Snippet: Modifications on original strain affecting melatonin sensing properties. A comparison of the dose–response between original biosensor strain and after increasing MTNR1A receptor number (yRB1002) ( A ), after increasing MTNR1A and adding one extra copy of the reporter cassette (yRB1012) ( B ) and after increasing MTNR1A and integrating reporter cassette multiple times (yRB1022) ( C ). Measurements are normalized sfGFP fluorescence levels and shown as mean ± SD of triplicates. Curve fitting and parameters were obtained using GraphPad Prism 9.5.1 variable slope (four parameter) nonlinear regression fit.

Article Snippet: Yeast biosensor strain yRB1012 was employed in this assay. sfGFP fluorescence for each sample was interpolated in the fitted calibration curve for melatonin using GraphPad Prism 9.5.1 standard curve interpolation (four parameter) tool.

Techniques: Comparison, Fluorescence

Journal: Scientific Reports

Article Title: Engineering a GPCR-based yeast biosensor for a highly sensitive melatonin detection from fermented beverages

doi: 10.1038/s41598-024-68633-y

Figure Lengend Snippet: Screening of 101 yeast strains. Yeast biosensor strain yRB1012 was employed in this assay. sfGFP fluorescence for each sample was interpolated in the fitted calibration curve for melatonin using GraphPad Prism 9.5.1 standard curve interpolation (four parameter) tool. Melatonin concentration values are shown as mean ± SD of triplicates. 17 of the 101 strains assayed were excluded from interpolation due to lack of enough signal.

Article Snippet: Yeast biosensor strain yRB1012 was employed in this assay. sfGFP fluorescence for each sample was interpolated in the fitted calibration curve for melatonin using GraphPad Prism 9.5.1 standard curve interpolation (four parameter) tool.

Techniques: Fluorescence, Concentration Assay