基于TTC染色法的高活力酵母细胞定量筛选  

A rapid and dynamic quantitative screening method based on TTC staining of high activity yeast cells

作　　者：徐俊[1] 雍晓雨[1] 费文斌[1] 周俊[1,2] 王舒雅[1,2] 陈怡露[1] 郑涛[1,2] 

机构地区：[1]南京工业大学生物与制药工程学院,江苏南京211816 [2]南京工业大学生物能源研究所,江苏南京211816

出　　处：《食品与发酵工业》2014年第7期1-5,共5页Food and Fermentation Industries

基　　金：国家重点基础研究发展计划(973计划)(2013CB733904;2011CBA00807;2013CB733502);国家高技术研究发展计划(863计划)(2012AA022106);江苏省自然科学基金(BK20130932)

摘　　要：在2,3,5-氯化三苯基四氮唑(TTC)染色法及96孔板基础上建立高活力与高产乙醇酵母细胞的同步定量筛选方法。选用体积分数为85%二甲基亚砜作为TTF提取剂,培养液(pH8.0)中TTC添加浓度为0.4%,以96孔板在0%、10%、14%的乙醇浓度下培养27株备选酵母菌株,通过酶标仪测定波长486 nm下TTF吸收值。结果显示TTF吸收值越高,酵母活性越高,并且通过比较酵母活性与其乙醇产量,得出两者在一定程度上呈正相关,即高活性酵母其乙醇产量也较高。共筛得6株具有较高细胞活性的菌株。此方法为高活性、高产乙醇酵母及其他微生物及植物细胞的活性的快速、大量检测提供了一条新途径。A quantitative screening method for yeast with high activity was established based on the TTC （Triphe- nyl- tetrazolium chloride） staining method and 96-well plate. The optimal test conditions were 85% （V/V） dimethyl sulfoxide, 0.4% TTC, pH 8, and testing wave length 486nm. This assay was performed in 96-well microtitre plate. Twenty seven strains were cultured in the 96-well microtitre plate with 0% , 10% , and 14% （V/V） ethanol. The TTF in cell was detected by microplate reader. Strains with higher TTF absorption values had higher ethanol producing ability. The research presented a novel rapid, high throughput method to screen yeast with high activity and high etha- nol yield. As results, 6 strains with high TTF absorption value were obtained. This screening method was also very useful to screen all kinds of microorganism and plant cells.

关 键 词：TTC 高活力酵母 乙醇产量 96孔板 定量筛选 

分 类 号：TQ920.6[轻工技术与工程—发酵工程]