Gustafsson Frida S, Whiteside Matthew D, Jiranek Vladimir, Durall Daniel M
Biology Department, The University of British Columbia Okanagan, 3333 University Way, Kelowna, BC V1V 1V7, Canada.
1] Biology Department, The University of British Columbia Okanagan, 3333 University Way, Kelowna, BC V1V 1V7, Canada [2] School of Agriculture, Food and Wine, The University of Adelaide, PMB1, Glen Osmond, SA 5064, Australia.
Sci Rep. 2014 Nov 10;4:6971. doi: 10.1038/srep06971.
Saccharomyces cerevisiae strains vary in their ability to develop and enhance sensory attributes of alcoholic beverages and are often found growing in mixed strain fermentations; however, quantifying individual strains is challenging due to quantification inaccuracies, low marker longevity, and compromised kinetics. We developed a fluorescent probe, consisting of glutathione molecules conjugated to a quantum dot (QD). Two S. cerevisiae strains were incubated with different coloured probes (QD attached to glutathione molecules, QD-GSH), fermented at multiple ratios, and quantified using confocal microscopy. The QD method was compared with a culture method using microsatellite DNA analysis (MS method). Probes were taken up by an ADP1 encoded transporter, transferred from mother cell to daughter cell, detectable in strains throughout fermentation, and were non-toxic. This resulted in a new quantification method that was more accurate and efficient than the MS method.
酿酒酵母菌株在发展和增强酒精饮料感官特性的能力上存在差异,并且经常在混合菌株发酵中生长;然而,由于定量不准确、标记物寿命短和动力学受损,对单个菌株进行定量具有挑战性。我们开发了一种荧光探针,它由与量子点(QD)偶联的谷胱甘肽分子组成。将两种酿酒酵母菌株与不同颜色的探针(连接到谷胱甘肽分子上的量子点,QD-GSH)一起孵育,以多种比例进行发酵,并使用共聚焦显微镜进行定量。将量子点方法与使用微卫星DNA分析的培养方法(MS方法)进行比较。探针被一种ADP1编码的转运蛋白摄取,从母细胞转移到子细胞,在整个发酵过程中在菌株中均可检测到,并且无毒。这产生了一种比MS方法更准确、更有效的新定量方法。