TTC比色法筛选高存活率的毕赤酵母突变株  

Screening of High Cell Viability Pichia pastoris Mutant by TTC Colorimetric Method

作　　者：刘胜男[1] 张燕[1] 刘泰瑜 许可[1] 张建国[1] 

机构地区：[1]上海理工大学食品科学与工程研究所,上海200093

出　　处：《化学与生物工程》2016年第10期18-22,共5页Chemistry & Bioengineering

基　　金：国家自然科学基金资助项目(21306112);上海市自然科学基金资助项目(13ZR1429100);上海高校青年教师培养资助计划项目(slg14037);教育部留学回国人员科研启动基金资助项目;上海理工大学研究生创新基金资助项目;微创励志创新基金资助项目(YS30808007)

摘　　要：采用TTC比色法筛选高存活率毕赤酵母NTG突变株,探讨了pH值、菌体浓度（OD600）、反应时间、细胞存活率等对TTC显色反应的影响,并与平板计数法进行比较。结果表明,在pH值为4.0~7.0或菌体浓度（OD600）低于50.00时,pH值、OD600均与1,3,5-三苯基甲臜（TPF）生成量（OD485）呈线性正相关关系;反应时间超过9h后可以保持稳定的TPF生成量;死细胞对TTC显色反应无干扰,在细胞存活率低于50%时,仍然可以采用TTC比色法测定细胞活性。表明,TTC比色法在一定范围内可以筛选高存活率毕赤酵母突变株,且操作简单省时,为筛选高存活率的微生物菌株提供了可靠技术。High cell viability Pichia pastoris mutant was screened by TTC colorimetric method.The effects of pH value,cell density（OD600）,reaction time,cell viability on TTC colorimetric reaction were investigated,and compared with the plate counting method.Results indicated that OD485 of 1,3,5-triphenylformazan（TPF）was proportional related to the pH value（4.0~7.0）or OD600（below 50.00）.And the production of TPF remained stable after reaction for 9h.Dead cells did not interfere with colorimetric reaction.So TTC colorimetric method could be used to detect cell activity even when the cell viability was bellow 50%.TTC colorimetric method could be applied in screening of high cell viability Pichia pastoris mutant within a certain range,and it was simple and time-saving.In summary,TTC colorimetric method provided a reliable approach for screening of high cell viability strain.

关 键 词：TTC比色法 毕赤酵母 存活率 优化 亚硝基胍 筛选 

分 类 号：Q815[生物学—生物工程]