Department of Food Science and Human Nutrition, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.
Enzyme Microb Technol. 2013 Aug 15;53(3):174-80. doi: 10.1016/j.enzmictec.2013.03.016. Epub 2013 Mar 29.
Protein expression patterns of an erythritol-producing yeast, Candida magnoliae, were analyzed to identify differentially expressed proteins in response to glucose perturbation. Specifically, wild type C. magnoliae was grown under high and low glucose conditions and the cells were harvested at both mid-exponential and erythritol production phases for proteomic studies. In order to analyze intracellular protein abundances from the harvested cells quantitatively, total intracellular proteins were extracted and applied to two-dimensional gel electrophoresis for separation and visualization of individual proteins. Among the proteins distributed in the range of pI 4-7 and molecular weight 29-97kDa, five osmo-responsive proteins were drastically changed in response to glucose perturbation. Hsp60 (Heat-shock protein 60), transaldolase and NADH:quinone oxidoreductase were down-regulated under the high glucose condition and Bro1 (BCK1-like Resistance to Osmotic shock) and Eno1 (enolase1) were up-regulated. These proteins are directly or indirectly related with cellular stress response. Importantly, protein expression patterns of Hsp60, Bro1 and Eno1 were strongly correlated with previous studies identifying the proteins perturbed by osmotic stress for other organisms including Saccharomyces cerevisiae.
为了鉴定在响应葡萄糖胁迫时差异表达的蛋白质，分析了产赤藓糖醇酵母 Candida magnoliae 的蛋白质表达模式。具体来说，在高葡萄糖和低葡萄糖条件下培养野生型 C. magnoliae，并在细胞生长的指数中期和赤藓糖醇生产阶段收获细胞以进行蛋白质组学研究。为了从收获的细胞中定量分析细胞内蛋白质丰度，提取了总细胞内蛋白质，并将其应用于二维凝胶电泳，以分离和可视化单个蛋白质。在等电点为 4-7 和分子量为 29-97 kDa 的范围内分布的蛋白质中，有 5 种与渗透压应激相关的蛋白质对葡萄糖胁迫有明显的变化。在高葡萄糖条件下，Hsp60（热休克蛋白 60）、转醛醇酶和 NADH：醌氧化还原酶下调，而 Bro1（BCK1 样耐渗透压冲击）和 Eno1（烯醇酶 1）上调。这些蛋白质直接或间接地与细胞应激反应有关。重要的是，Hsp60、Bro1 和 Eno1 的蛋白质表达模式与之前的研究强烈相关，该研究确定了包括酿酒酵母在内的其他生物体中受渗透压应激影响的蛋白质。
