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Table 3 Total functional yields from TM6* producing two human GPCRs are at least quadruple that from the corresponding wild-type transformants

From: Increasing cell biomass in Saccharomyces cerevisiae increases recombinant protein yield: the use of a respiratory strain as a microbial cell factory

Transformants Duration of culture (h) Specific growth rate (h-1) Dry cell weight yield (g g-1 Glc) Harvest wet cell weight (g) Total membrane protein yield (mg) Bmax (pmol mg-1) Kd (nM)
WT CNR2 16 0.3 (0.03) 0.12 (0.02) 5.0 5.1 (0.48) 22.02 (4.00) 39.75 (12.96)
WT A2aR 16 0.3 (0.03) 0.12 (0.02) 4.0 5.8 (0.19) 5.12 (0.59) 5.52 (1.68)
TM6* CNR2 25 0.2 (0.02) 0.32 (0.02) 9.0 23.3 (1.00) 17.02 (2.20) 30.81 (7.71)
TM6* A2aR 25 0.2 (0.02) 0.32 (0.02) 10 29.4 (0.98) 5.16 (0.61) 6.26 (1.86)
  1. The human GPCRs, A2aR and CNR2, were produced in the wild-type S. cerevisiae strain, BY4741 (WT), and the respiratory strain, TM6*. Cells were harvested at the time points indicated for preparation of membranes for functional analysis, as described in the Methods section. The mean growth rate (h-1) is calculated for the exponential growth phase (n = 2). The mean dry cell weight yield is reported with respect to the glucose consumed (n = 2). Harvest wet cell weight is reported in g, and the mean total membrane protein yield from these cells is reported (n = 3). Bmax and Kd values were calculated from the curves shown in Figure 5 (n = 3). In all cases the standard error of the mean is given in parentheses.