The BioTechnology Institute, University of Minnesota Twin Cities, St. Paul, MN, 55108, USA.
The BioTechnology Institute, University of Minnesota Twin Cities, St. Paul, MN, 55108, USA; Department of Civil, Environmental, and Geo-Engineering, University of Minnesota Twin Cities, Minneapolis, MN, 55455, USA.
Water Res. 2021 Oct 15;205:117696. doi: 10.1016/j.watres.2021.117696. Epub 2021 Sep 24.
In order to ensure stable performance of engineered biotechnologies that rely on mixed microbial community systems, it is important to identify process-specific microbial traits and study their in-situ activity and responses to changing environmental conditions and system operational parameters. We used BioOrthogonal Non-Canonical Amino acid Tagging (BONCAT) in combination with Fluorescence-Activated Cell Sorting (FACS) and 16S rRNA gene amplicon sequencing to identify translationally active cells in activated sludge. We found that only a subset of the activated sludge microbiome is translationally active during the aerobic treatment phase of a full-scale sequencing batch reactor designed to enhance biological phosphorus removal from municipal wastewater. Relative abundance of amplicon sequence variants was not a reliable predictor of species activity. BONCAT-positive and -negative cells revealed a broad range of population-wide and taxa-specific translational heterogeneity. BONCAT-FACS in combination with amplicon sequencing can provide new insights into the ecophysiology of highly dynamic microbiomes in activated sludge systems.
为了确保依赖混合微生物群落系统的工程生物技术具有稳定的性能，识别特定于工艺的微生物特性并研究其原位活性以及对环境条件变化和系统运行参数的响应非常重要。我们使用生物正交非天然氨基酸标记（BONCAT）结合荧光激活细胞分选（FACS）和 16S rRNA 基因扩增子测序来鉴定活性污泥中的翻译活性细胞。我们发现，在设计用于从城市废水中增强生物除磷的全规模序批式反应器的好氧处理阶段，只有活性污泥微生物组的一部分具有翻译活性。扩增子序列变体的相对丰度不是物种活性的可靠预测因子。BONCAT 阳性和阴性细胞揭示了广泛的全种群和分类特异性翻译异质性。BONCAT-FACS 结合扩增子测序可以为活性污泥系统中高度动态微生物组的生态生理学提供新的见解。
