Nishino Tomohiko, Matsuda Yusuke, Yamazaki Yuna
School of Bioscience and Biotechnology, Tokyo University of Technology, 1404 Katakura, Hachioji, Tokyo, 192-0982, Japan.
Heliyon. 2018 Apr 11;4(4):e00597. doi: 10.1016/j.heliyon.2018.e00597. eCollection 2018 Apr.
Probiotics are live microorganisms that provide health benefits to humans. Some lactic acid bacteria (LAB) are probiotic organisms used in the production of fermented foods, such as yogurt, cheese, and pickles. Given their widespread consumption, it is important to understand the physiological state of LAB in foods such as yogurt. However, this analysis is complicated, as it is difficult to separate the LAB from milk components such as solid curds, which prevent cell separation by dilution or centrifugation. In this study, we successfully separated viable LAB from yogurt by density gradient centrifugation. The recovery rate was >90 %, and separation was performed until the stationary phase. Recovered cells were observable by microscopy, meaning that morphological changes and cell viability could be directly detected at the single-cell level. The results indicate that viable LAB can be easily purified from fermented milk. We expect that this method will be a useful tool for the analysis of various aspects of probiotic cells, including their enzyme activity and protein expression.
益生菌是对人类健康有益的活微生物。一些乳酸菌(LAB)是用于生产发酵食品(如酸奶、奶酪和泡菜)的益生菌。鉴于它们的广泛消费,了解酸奶等食品中乳酸菌的生理状态很重要。然而,这种分析很复杂,因为很难将乳酸菌与牛奶成分(如固体凝乳)分离,这些成分会阻止通过稀释或离心进行细胞分离。在本研究中,我们通过密度梯度离心成功地从酸奶中分离出有活力的乳酸菌。回收率>90%,并且分离一直进行到稳定期。通过显微镜可以观察到回收的细胞,这意味着可以在单细胞水平直接检测形态变化和细胞活力。结果表明,有活力的乳酸菌可以很容易地从发酵乳中纯化出来。我们预计这种方法将成为分析益生菌细胞各个方面(包括其酶活性和蛋白质表达)的有用工具。