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Fig. 2 | Microbial Cell Factories

Fig. 2

From: Development of a yeast whole-cell biocatalyst for MHET conversion into terephthalic acid and ethylene glycol

Fig. 2

MHETase display constructs are efficiently expressed at minimal fitness cost. A GFP calibration standards for measuring abundance of MHETase chimeras in molecules per cell. GFP-fusion strains spanning the range of molecules per cell were selected and GFP fluorescence was measured. The regression analysis line and equation are indicated. Bars indicate standard deviation; n ≥ 7. B Abundance of the indicated surface display chimeras with (orange) or without (green) MHETase. Abundance was determined using GFP fluorescence after induction with doxycycline for 4 h and converted to molecules per cell using the equation in A. Theoretical MHETase molarity was inferred from the molecule/cell data for a cell density of 108 cells/mL (right y-axis). Horizontal bars indicate the means of the replicates. Asterisks indicate p-values ≤ 0.05 (unpaired Student’s t-test; n = 7). Intracellular MHETase (intra-M) and secreted MHETase (secreted-M) are indicated. C. Fitness of cells expressing the surface display chimeras. Cells expressing the indicated chimeras were grown in presence of doxycycline in YPD medium for 24 h. Fitness is expressed as a ratio of the growth rate of each strain to that of the wild-type. Horizontal bars indicate the means of the replicates. Asterisks indicate p-values ≤ 0.05 (unpaired Student’s t-test; n = 4)

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