Monitoring the intracellular pH of Zygosaccharomyces bailii by green fluorescent protein
Monitoring the intracellular pH of Zygosaccharomyces bailii by green fluorescent protein
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DOI：
10.1016/j.ijfoodmicro.2012.03.028
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发表时间：
2012-06-01
影响因子：
5.4
通讯作者：
Devlieghere, F.
中科院分区：
文献类型：
--
作者：
Dang, T. D. T.;De Maeseneire, S. L.;Devlieghere, F.
众所周知，细胞内pH（pH（i））在细胞生理学中起着至关重要的作用，并且pH（i）稳态对于正常的细胞功能是必不可少的。因此，需要知道在细胞生命周期期间或在各种生长条件下的pH（i）。已经开发了不同的测量pH（i）的方法，并且在这些方法中，使用pH敏感的绿色荧光蛋白（GFP）作为pH（i）指示剂是一种有前途的技术。通过使用这种方法，不仅可以获得更准确的pH（i）结果，而且可以对pH（i）进行长期实验。在这项研究中，野生型接合酵母贝利。用编码pH敏感性GFP（即pH1uorin）的质粒转化臭名昭著的食品腐败酵母，使得能够基于细胞荧光信号确定酵母的pH（i）。转化后，在22 ℃下在四种不同的酸性条件下研究酵母的生长和pHi 26天。从实验结果中，转化效果得到验证，酵母生长和pH（i）之间的良好的相关性被注意到。特别地，观察到酵母具有在指数期期间耐受显著的pH（i）下降和随后在稳定期中耐受pH（i）恢复的能力，这可能是酵母异常的耐酸性的基础。这是首次将基于GFP的pH（i）测量方法应用于Z。bailii，并且在如此长的时间段（26天）期间监测酵母的pH（i）。通过对该菌株的进一步研究，可以进一步了解该菌株的生理特性和耐酸性机制。(C)2012爱思唯尔有限公司版权所有。
It is generally known that intracellular pH (pH(i)) plays a vital role in cell physiology and that pH(i) homeostasis is essential for normal cellular functions. Therefore, it is desirable to know the pH(i) during cell life cycle or under various growth conditions. Different methods to measure pH(i) have been developed and among these methods, the use of pH-sensitive green fluorescent protein (GFP) as a pH(i) indicator is a promising technique. By using this approach, not only can more accurate pH(i) results be obtained but also long-term experiments on pH(i) can be performed. In this study, the wild type Zygosaccharomyces bailii. a notorious food spoilage yeast, was transformed with a plasmid encoding a pH-sensitive GFP (i.e. pHIuorin), enabling the pH(i) of the yeast to be determined based on cellular fluorescent signals. After the transformation, growth and pHi of the yeast were investigated in four different acidic conditions at 22 degrees C during 26 days. From the experimental results, the transformation effectiveness was verified and a good correlation between yeast growth and pH(i) was noticed. Particularly, it was observed that the yeast has an ability to tolerate a significant pH(i) drop during exponential phase and a subsequent pH(i) recovery in stationary phase, which may underlie the exceptional acid resistance of the yeast. This was the first time that a GFP-based approach for pH(i) measurement was applied in Z. bailii and that the pH(i) of the yeast was monitored during such a long period (26 days). It can be expected that greater understanding of the physiological properties and mechanisms behind the special acid resistance of the yeast will be obtained from further studies on this new yeast strain. (C) 2012 Elsevier B.V. All rights reserved.