| 毕赤酵母发酵过程胞内活性氧的定量分析 |
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| 引用本文： | 肖安风,孙亮先,倪辉,蔡慧农.毕赤酵母发酵过程胞内活性氧的定量分析[J].泉州师范学院学报,2009,27(4):87-90. |
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| 作者姓名： | 肖安风 孙亮先 倪辉 蔡慧农 |
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| 作者单位： | 1. 集美大学,生物工程学院,福建,厦门,361021;厦门市食品生物工程技术研究中心,福建,厦门,361021
2. 泉州师范学院,模式生物研究中心,福建,泉州,362000 |
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| 摘 要： | 利用流式细胞术对毕赤酵母发酵过程中胞内活性氧（ROS）的变化进行定量检测和分析，建立发酵过程单位细胞胞内ROS的相对含量以及单位体积细胞胞内ROS相对含量的计算方法，分析ROS积累对毕赤酵母细胞活力的影响,研究结果表明：在甲醇流加阶段，单位细胞的DCF（2′，7′-二氯荧光黄）染色平均荧光强度随着时间的增加而增加，即单位细胞胞内ROS的含量在甲醇流加阶段一直增加，细胞内产生持续的氧化压力，导致部分细胞死亡；单位体积酵母细胞的胞内ROS含量在甲醇流加阶段保持在一个相对稳定的范围。
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| 关 键 词： | 毕赤酵母 活性氧 流式细胞术 细胞活力 定量分析 |
Quantitative Analysis of Intracellular Reactive Oxygen Species in Pichia Pastoris Fermentation with Flow Cytometry |
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| XIAO An-feng,SUN Liang-xian,NI Hui,CAI Hui-nong.Quantitative Analysis of Intracellular Reactive Oxygen Species in Pichia Pastoris Fermentation with Flow Cytometry[J].Journal of Quanzhou Normal College,2009,27(4):87-90. |
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| Authors: | XIAO An-feng SUN Liang-xian NI Hui CAI Hui-nong |
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| Institution: | 1. College of Bioengineering,Jimei University,Xiamen 361021 ,Chinal2. Model Organism Research Centre, Quanzhou Normal College, Fujian 36200, China3. Research Center of Food Biotechnology, Xiamen 361021 ,China) |
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| Abstract: | Intracellular reactive oxygen species （ROS） were detected by flow cytometry in Pichia pastoris fermentation. Through the analysis of fluorescence staining signal,ways of calculating relative intracellular ROS level per cell and per cell volume were founded and used to analyze the correlation of ROS and cell viability. In this study, the stage of cell damage during fermentation was correlated with the accumulation of ROS. At the glycerol batch and fed-batch phase,little ROS could he detected and cell viability reached almost 100%. At the methanol fed-batch phase, DCF （2′, 7′-dichlorfluorescein） staining intensity of single cell increased along with time passing, which meant that intracellular ROS level of single cell was increasing at this phase producing everlasting oxygen pressure to yeast cell. As a result,some cells lost viability. In the mean time,intracellular ROS level of unit cell volume kept in a stable level at the methanol fed-batch phase. |
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| Keywords: | Pichia pastoris reactive oxygen species flow cytometry cell viability quantitative analysis |
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