K. phaffii strain BSYBG10 with multiple inducers, sampled after 3h
A commercially available WT P. pastoris strain BSYBG10 (bisy GmbH, Hofstaetten/Raab, Austria) was cultivated in 25 mL minimal media (BM, 200 mM potassium phosphate pH 6, 1.34 % YNB, 4 x 10-5 % biotin) with 0.25% glycerol as carbon source in 250 mL baffled shake flasks for 9 hours (28?C, 130 rpm, 80% humidity) until the initial glycerol was depleted. Afterwards, two glycerol FeedBeads were added, followed by the addition of 25 mL fresh minimal media supplemented with different inductors (xylitol, galactose, xylose, sorbitol) to an end concentration of 0.25% in the media. After three hours, samples were taken and centrifuged at 5000*g. The cell pellet was frozen at -80?C and sent for RNA isolation and sequencing at BioGrammatics Inc, Carlbad, California US. Total RNA extraction, as well as quality control and library preparation were performed according to BioGrammatics standards. Poly-A enriched total RNA was sequenced as 50 bp single reads on an Illumina sequencer.