工业模式微生物膜有序性的活细胞定量分析 <sup>*</sup>

doi:10.13523/j.cb.2008140

中国生物工程杂志

2021, Vol. 41

Issue (1): 20-29 DOI: 10.13523/j.cb.2008140

研究报告

工业模式微生物膜有序性的活细胞定量分析 ^*

吕雪芹,金柯,刘家恒,崔世修,李江华,堵国成,刘龙(

)

江南大学未来食品科技中心化学与生物技术教育部重点实验室无锡 214122

Quantitative Analysis of Membrane Ordering of Living Industrial Model Microorganisms

LV Xue-qin,JIN Ke,LIU Jia-heng,CUI Shi-xiu,LI Jiang-hua,DU Guo-cheng,LIU Long(

)

Science Center for Future Foods, Jiangnan University, Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Wuxi 214122, China

全文: PDF(61192 KB) HTML

摘要：

作为一种相位敏感的荧光探针,Di-4-ANEPPDHQ可以特异性标记膜的有序相和无序相,在理论上可以对细胞膜的有序性进行定量成像。通过将Di-4-ANEPPDHQ和激光扫描共聚焦显微术相结合,对多种具有代表性的工业模式微生物进行了有序相和无序相活细胞成像,结合极性归一化数值的统计比较,最终实现对上述工业模式微生物细胞膜有序性的定量分析,为细胞膜工程提供了一种直观且快速的活细胞检测方法。

关键词： 工业模式微生物; Di-4-ANEPPDHQ; 膜有序性; 活细胞成像

Abstract:

As a phase sensitive fluorescent probe, Di-4-ANEPPDHQ can specifically label the ordered phase and disordered phase of membrane. Therefore, in theory, the probe can be used to quantitatively image the order of cell membrane. By combining Di-4-ANEPPDHQ and laser scanning confocal microscopy, the ordered phase and disordered phase live-cell imaging of a variety of representative industrial model microorganisms were carried out. Combined with the statistical comparison of polarity normalization values, the quantitative analysis of the cell membrane ordering of the above industrial model microorganisms is finally realized. The above study provides an intuitive and rapid detection method of living cells for cell membrane engineering.

Key words: Industrial model microorganisms Di-4-ANEPPDHQ Membrane ordering Live-cell imaging

收稿日期: 2020-08-27 出版日期: 2021-02-09

ZTFLH:

Q819

基金资助: * 国家自然科学基金(21808084);国家重点研发计划(2018YFA0900300)

通讯作者: 刘龙 E-mail: liulong@jiangnan.edu.cn

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	吕雪芹
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引用本文:

吕雪芹, 金柯, 刘家恒, 崔世修, 李江华, 堵国成, 刘龙. 工业模式微生物膜有序性的活细胞定量分析 ^*[J]. 中国生物工程杂志, 2021, 41(1): 20-29.

LV Xue-qin, JIN Ke, LIU Jia-heng, CUI Shi-xiu, LI Jiang-hua, DU Guo-cheng, LIU Long. Quantitative Analysis of Membrane Ordering of Living Industrial Model Microorganisms. China Biotechnology, 2021, 41(1): 20-29.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.2008140 或 https://manu60.magtech.com.cn/biotech/CN/Y2021/V41/I1/20

图1 Di-4-ANEPPDHQ分子结构式

图2 酿酒酵母Di-4-ANEPPDHQ的发射光谱分析

图3 酿酒酵母膜有序度分析

图4 大肠杆菌DI-4-ANEPPDHQ染色效果分析

图5 枯草芽孢杆菌中DI-4-ANEPPDHQ的发射光谱和膜有序度分析

图6 巨大芽孢杆菌和谷氨酸棒杆菌中Di-4-ANEPPDHQ发射光谱的分析和比较

图7 三种革兰氏阳性菌之间的膜有序度分析和比较

图8 BS-168和BS-MK7菌株中MK-7的产量

图9 不同枯草芽孢改造菌株中膜有序度分析

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