酵母中性脂快速检测及积累动态分析  

Rapid Quantitation and Dynamic Accumulation Analysis of Neutral Lipids in Yeast

作　　者：杜秀秀[1] 房志家[1] 陈忠翔[1] 匡鑫[1] 黄志伟[1] 

机构地区：[1]东华大学化学化工与生物工程学院,上海201620

出　　处：《生物技术通报》2014年第8期208-214,共7页Biotechnology Bulletin

基　　金：国家自然科学基金项目(31100549);中央高校基本科研业务费专项资金项目(22320143-03)

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。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 total neutral lipid levels did not change;while triglyceride and total neutral lipids contents were significantly accumulated in TGL knockout mutant. Finally, the dynamic changes of yeast neutral lipids content were successfully monitored with the change of growth phase by this method, suggesting that the neutral lipids were obviously accumulated after exponential phase.

关 键 词：出芽酵母 中性脂 亲脂染料 脂肪代谢调控 

分 类 号：Q936[生物学—微生物学]