e. storage proteins from barley, and many of the trypsin/α-amylase inhibitors from barley, vanish during the process of making beer (wort boiling and fermentation) and only half of the proteins identified in barley grain were also present in beer. Other studies used two-dimensional gel electrophoresis (2-DE) to discover proteins involved

in head foam and beer haze formation [13–16] and Z-IETD-FMK nmr the influence of malt modification and processing [6, 14]. Proteins derived from brewer’s yeast have also been identified in beer, although the range of identified proteins vary from 2–4 proteins [8, 17] to 31 proteins [5] and 40 protein fragments [4]. The origin of the identified proteins also vary from proteins localized in the cytosol, such as enolase and triosephosphate isomerase, to proteins like Swc4 and Uth1 that are associated to the cell wall [4, 5, 8]. One common feature

for all beer proteome studies, so far, is that commercial beers have been used where no information CP 690550 on raw materials, choice of brewer’s yeast strain, or fermentation conditions have been given. In this study, we used two ale brewer’s yeast strains, differing in their ability to consume fermentable sugars, for brewing beer under controlled conditions to determine the protein changes caused by fermentation, and to explore if there are any yeast strain dependent changes of the beer proteome. Methods Yeast strains and media The yeast strains (WLP001 and KVL011) used in this study were ale brewer’s yeast strains, belonging to the species Saccharomyces cerevisiae, obtained from White Labs (WL, San Diego, California, USA) and our own collection (KVL) at the Department of Food Science, Food Microbiology, University of Copenhagen, respectively. Yeast strains were grown in 0.3% malt extract, 0.3% yeast extract, 0.5% peptone, 1% glucose, pH5.6 (MYGP) or in standard

hopped wort (13° Plato) from Skands Brewery (Skands, Brøndby, Denmark). Beer fermentation Aerobic propagation of yeast was started from a single colony on a MYGP-agar plate in 10 ml MYGP, in duplicate. After incubation at 20°C for 24 h, the yeast suspensions were transferred to 100 ml MYGP in 250 ml Erlenmeyer flasks with aeration at 200 rpm. Yeast suspensions were transferred after two days at 20°C to 400 ml double concentrated MYGP and incubated for 24 h at 20°C. Yeast cells were harvested (3000 g, 10 min, 20°C) and Sinomenine inoculated at 7 × 106 cells/ml in 2 litres of wort saturated with air. Fermentations were carried out in biological duplicates in 2.5-liters European Brewing Convention (EBC) tubes at 18°C for 155 hours. To monitor the fermentation, samples of culture broth were collected aseptically twice on a daily basis from the top of the EBC-tubes for 155 hours. Yeast growth was followed by LY2835219 measuring the optical density at 600 nm (OD600)(UV-1800; Shimadzu Scientific Instruments) and pH (pHM220; Radiometer Analytical SAS). Sugar and ethanol determination Samples were filtrated using a 0.