The field of systems biology is often held back by difficulties in obtaining comprehensive, highquality, quantitative data sets. In this paper, we undertook an interlaboratory effort to generate such a data set for a very large number of cellular components in the yeast Saccharomyces cerevisiae, a widely used model organism that is also used in the production of fuels, chemicals, food ingredients and pharmaceuticals. With the current focus on biofuels and sustainability, there is much interest in harnessing this species as a general cell factory. In this study, we characterized two yeast strains, under two standard growth conditions. We ensured the high quality of the experimental data by evaluating a wide range of sampling and analytical techniques. Here we show significant differences in the maximum specific growth rate and biomass yield between the two strains. on the basis of the integrated analysis of the highthroughput data, we hypothesize that differences in phenotype are due to differences in protein metabolism.

We here present simple and rapid methods for fast screening of yeast lipids in Saccharomyces cerevisiae. First we introduced a microwave-assisted technique for fast lipid extraction that allows the extraction of lipids within 10 min. The new method enhances extraction rate by 27 times, while maintaining product yields comparable to conventional methods (n = 14, P > 0.05). The recovery (n = 3) from spiking of synthetic standards were 92 ± 6% for cholesterol, 95 ± 4% for triacylglycerol, and 92 ± 4% for free fatty acids. Additionally, the new extraction method combines cell disruption and extraction in one step, and the approach, therefore, not only greatly simplifies sample handling but also reduces analysis time and minimizes sample loss during sample preparation. Second, we developed a chromatographic separation that allowed separation of neutral and polar lipids from the extracted samples within a single run. The separation was performed based on a three gradient solvent system combined with hydrophilic interaction liquid chromatography-HPLC followed by detection using a charged aerosol detector. The method was shown to be highly reproducible in terms of retention time of the analytes (intraday; 0.002-0.034% RSD; n = 10, interday; 0.04-1.35% RSD; n = 5) and peak area (intraday; 0.63-6% RSD; n = 10, interday; 4-12% RSD; n = 5).

Nutrient sensing and coordination of metabolic pathways are crucial functions for living cells. A combined analysis of the yeast transcriptome, phosphoproteome and metabolome is used to investigate the interactions between the Snf1 and TORC1 pathways under nutrient-limited conditions.

We present a fast and accurate method for preparation of fatty acid methyl esters (FAMEs) using microwave-assisted derivatization of fatty acids present in yeast samples. The esterification of free/bound fatty acids to FAMEs was completed within 5 min, which is 24 times faster than with conventional heating methods. The developed method was validated in two ways: (1) through comparison with a conventional method (hot plate) and (2) through validation with the standard reference material (SRM) 3275-2 omega-3 and omega-6 fatty acids in fish oil (from the Nation Institute of Standards and Technology, USA). There were no significant differences (P>0.05) in yields of FAMEs with both validations. By performing a simple modification of closed-vessel microwave heating, it was possible to carry out the esterification in Pyrex glass tubes kept inside the closed vessel. Hereby, we are able to increase the number of sample preparations to several hundred samples per day as the time for preparation of reused vessels was eliminated. Pretreated cell disruption steps are not required, since the direct FAME preparation provides equally quantitative results. The new microwave-assisted derivatization method facilitates the preparation of FAMEs directly from yeast cells, but the method is likely to also be applicable for other biological samples.

Several reports have shown antimicrobial and anticancer activities of mucous glycoproteins extracted from the giant African snail Achatina fulica. Anticancer properties of the snail mucous peptides remain incompletely revealed. The aim of this study was to predict anticancer peptides from A. fulica mucus. Two of HPLC-separated mucous fractions (F2 and F5) showed in vitro cytotoxicity against the breast cancer cell line (MCF-7) and normal epithelium cell line (Vero). According to the mass spectrometric analysis, 404 and 424 peptides from the F2 and F5 fractions were identified. Our comprehensive bioinformatics workflow predicted 16 putative cationic and amphipathic anticancer peptides with diverse structures from these two peptidome data. These peptides would be promising molecules for new anti-breast cancer drug development.