15 research outputs found

    Timantinkaltaisen hiilipinnoitteen mekaaninen käyttäytyminen mikrosysteemeissä

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    The field of systems biology is often held back by difficulties in obtaining comprehensive, high-quality, 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 high-throughput data, we hypothesize that differences in phenotype are due to differences in protein metabolism

    Genome-Scale Metabolic Modeling from Yeast to Human Cell Models of Complex Diseases: Latest Advances and Challenges

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    Genome-scale metabolic models (GEMs) are mathematical models that enable systematic analysis of metabolism. This modeling concept has been applied to study the metabolism of many organisms including the eukaryal model organism, the yeast Saccharomyces cerevisiae, that also serves as an important cell factory for production of fuels and chemicals. With the application of yeast GEMs, our knowledge of metabolism is increasing. Therefore, GEMs have also been used for modeling human cells to study metabolic diseases. Here we introduce the concept of GEMs and provide a protocol for reconstructing GEMs. Besides, we show the historic development of yeast GEMs and their applications. Also, we review human GEMs as well as their uses in the studies of complex diseases

    Rapid Quantification of Yeast Lipid using Microwave-Assisted Total Lipid Extraction and HPLC-CAD

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    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)
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