organism: Saccharomyces cerevisiae S288C
description: Protocols: Saccharomyces cerevisiae W303 was used throughout the study. Three replicate batch experiments were carried out on ethanol as the only carbon source. Osmostress (final concentration of 0.4 M NaCl) was applied when biomass concentration had increased higher then 1.2 g L-1. Samples for cell dry weight, extracellular metabolites, trehalose and transcriptome analysis were collected just before stress application and 15, 30, 60 and 90 min after. Culture were grown at 30oC in 1.2 L bioreactors (DASGIP, Germany) with working volume of 0.75 L. The temperature, agitation, gassing, pH, and composition of the off gas were monitored and/or controlled using a DasGip monitoring and control system (DasGip, Germany). The stirrer speed was set to 400 rpm, and the aeration rate was 1 volume of gas per volume of fermentation broth per minute (vvm).The pH was controlled at 5.00±0.05 with 2 M KOH and the dissolved oxygen was kept above 30%. The CO2 emission (and residual O2) was monitored from the exhaust gas using the off-gas analyzer GA4 (DASGIP, Germany) , based on zirconium dioxide and a two-beam infrared sensor. Cultivation medium contained 40 g of ethanol, 15 g of (NH4)2SO4, 3 g of KH2PO4, 1.5 g of MgSO4 x 7 H2O, 1 mL of vitamin solution (Usaite et al, 2008), 1 mL of trace metal solution (Usaite et al, 2008), and 50 µL of Antiform 204 (Sigma-Aldrich, USA). Sampling for transcriptome analysis was carried out as follows: biomass was withdrawn from bioreactors into pre-cooled Eppendorf tubes, centrifuged for 22 sec at 4oC at 14 000 rpm and biomass was snap-freezed after decantating the supernatant. The whole sampling process took less than 45 sec. Samples were stored at -80oC. The Illumina TruSeq sample preparation kit v2, with poly-A selection, was used to prepare RNA samples for sequencing.
attributes.tag: ENA first public
attributes.value: 2017-02-27
attributes.tag: ENA last update
attributes.value: 2017-01-10
attributes.tag: External Id
attributes.value: SAMEA45291418
attributes.tag: INSDC center alias
attributes.value: Chalmers University of Technology, Department of Biology and Biological Engineering
attributes.tag: INSDC center name
attributes.value: Chalmers University of Technology, Department of Biology and Biological Engineering
attributes.tag: INSDC first public
attributes.value: 2017-02-27T17:01:50Z
attributes.tag: INSDC last update
attributes.value: 2017-01-10T09:20:01Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-5313:Sample 15
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: growth condition
attributes.value: batch growth on ethanol
attributes.tag: sample name
attributes.value: E-MTAB-5313:Sample 15
attributes.tag: strain
attributes.value: W303