Fig. 3
OPP-labelling assays can differentiate conditions and strains with altered translational activity. Assays were done using 0.3 g L− 1 OPP and 1.5 g L−1Imipramine. GlucLim denotes cultivation under limited glucose and GlucExc denotes cultivation under excess glucose, both in minimal media (ASMv6). All shown values were subtracted by the corresponding no-OPP control. Growth rates were determined by OD600 measurements in triplicate 1.5 h before and after sampling took place. a Relative translation activities of K. phaffii CBS2612 cells grown on different carbon sources. Growth rates were measured by OD600. In the MeOH 1 measurement, cells received 1% methanol 3 h before the assay. In MeOH 2, 1% methanol was given 3 h and another time 1.5 h before the sampling. Shown values were normalised to the glucose-excess condition fluorescence signals. All samples were measured in quadruplicate and a no-OPP control was made for each feed strategy in duplicate. b Translation activity of K. phaffii gcn2Δ relative to its unmodified parent. Cultivation was done in minimal media with glucose-limit to obtain equal growth rates. c Translation activities of GS115 PGAPHis + HAC1 relative to the empty vector control, GS115His+, under different cultivation conditions. Measurements were done in quadruplicate