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首页> 外文期刊>Journal of industrial microbiology & biotechnology >Monitoring growth phase-related changes in phosphatidylcholine-specific phospholipase C production, adhesion properties and physiology of Bacillus cereus vegetative cells
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Monitoring growth phase-related changes in phosphatidylcholine-specific phospholipase C production, adhesion properties and physiology of Bacillus cereus vegetative cells

机译：监测蜡状芽孢杆菌营养细胞的磷脂酰胆碱特异性磷脂酶C产生，粘附特性和生理生长期的相关变化

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摘要

The physiological status and metabolic heterogeneity of Bacillus cereus cells within a culture during an 8-h batch fermentation process was measured using flow cytometry (FCM). Concurrently, production of the toxin, PC-PLC, and the extent of cell adhesion of live and dead cells were monitored using novel fluorescent assays. Flow cytometry analysis detected growth phase-related changes in the physiological profiles of cells over the course of the fermentation, with variation in the percentage of cells displaying membrane damage and intracellular esterase and redox activities. As the exponential phase proceeded, populations became more uniform in terms of protein content as measured using FCM in tandem with a cell tracking dye, with the majority of cells becoming membrane intact, esterase positive and redox active. PC-PLC activity appeared strongly related to cell density. Permeabilisation of cells was accompanied by a loss in adherent properties, while 25-100% of cells with intracellular esterase activity possessed adhesion properties. Cells in late exponential phase appeared to have reduced adherence properties compared to cells in early exponential or lag phase. As well as demonstrating the utility of FCM for measuring heterogeneity in terms of cell physiological status throughout the course of batch cultures, the methods utilised in this study could be used to relate processes such as toxin production or cell adhesion to cell physiological state.
机译：使用流式细胞仪（FCM）测量了8小时分批发酵过程中培养物中蜡状芽孢杆菌细胞的生理状态和代谢异质性。同时，使用新型荧光测定法监测毒素，PC-PLC的产生以及活细胞和死细胞的细胞粘附程度。流式细胞仪分析检测到了发酵过程中细胞的生理特性与生长期相关的变化，其中细胞百分比的变化显示出膜损伤以及细胞内酯酶和氧化还原活性。随着指数期的进行，使用FCM与细胞跟踪染料串联测量时，种群的蛋白质含量变得更加均匀，大多数细胞变得完整，酯酶呈阳性，氧化还原活性。 PC-PLC活性似乎与细胞密度密切相关。细胞的通透性伴随着粘附特性的丧失，而具有细胞内酯酶活性的细胞中有25-100％具有粘附特性。与早期指数或滞后阶段的细胞相比，处于指数后期的细胞似乎具有降低的粘附特性。除了证明FCM在整个分批培养过程中用于测量细胞生理状态异质性的实用性外，本研究中使用的方法还可用于将毒素产生或细胞黏附等过程与细胞生理状态联系起来。

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