Table 3

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

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)

B_max (pmol mg^-1)

K_d (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 A_2aR

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* A_2aR

25

0.2 (0.02)

0.32 (0.02)

10

29.4 (0.98)

5.16 (0.61)

6.26 (1.86)

The human GPCRs, A_2aR 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). B_max and K_d 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.

Ferndahl et al. Microbial Cell Factories 2010 9:47   doi:10.1186/1475-2859-9-47