酵母中性脂快速检测及其积累动态分析.pdfVIP

生物技术通报 ·研究报告· BIOTECHNOLOGY BULLETIN 年第 期 2014 8 酵母中性脂快速检测及积累动态分析 杜秀秀  房志家  陈忠翔  匡鑫  黄志伟 （东华大学化学化工与生物工程学院，上海 201620） 摘 要 ： 以油红O、 苏丹黑B 为染料，结合酸或碱热破壁萃取，建立了通过比色快速检测酵母中性脂含量的方法。其中油 红O 萃取效果较好，比色结果更加准确，苏丹黑B 也可以作为一种替代的中性脂含量检测的染料。并用该方法初步研究了两种中 性脂肪甘油三酯代谢调控的关键基因磷脂酸磷酸酶 （PAH1）和甘油三酯脂酶 （triglyceride lipase，TGL）敲除突变体中性脂的变化。 结果显示，前者甘油三酯显著降低，但总中性脂肪含量不变，后者甘油三酯及总中性脂肪含量均明显积累 ；同时，用该方法监测 了酵母中性脂含量随生长周期的动态变化，表明指数生长期后有明显的积累。 关键词 ： 出芽酵母 中性脂 亲脂染料 脂肪代谢调控 Rapid Quantitation and Dynamic Accumulation Analysis of Neutral Lipids in Yeast Du Xiuxiu Fang Zhijia Chen Zhongxiang Kuang Xin Huang Zhiwei （ ， ， 201620 ） College of Chemistry Chemical Engineering and Biotechnology Donghua University Shanghai Abstract:  A colorimetric method for rapidly detecting neutral lipid content in yeast was developed, which was based on the staining of yeast cells by lipophilic dyes Oil red O or Sudan Black B and extraction with ethanol after cell rupturing using the method of combining hydrochloric acid or sodium hydroxide with heating. The extraction of Oil red O is better and the colorimetric results were more accurate, while Sudan Black B also can be used as an alternative dye for estimating neutral lipid content. Furthermore, the changes of neutral lipids in yeast mutated phospholipid fatty acid phosphatase（PAH1）and triglyceride lipase（TGL）, which are known as two key genes to regulate triglyceride metabolism, were studied using this method. Disruption of PAH1 resulted in a remarkable decrease in triglyceride content, although tota