Herbst, F.-A. | Search bioRxiv

bioRxiv2019

The Proteome of Tetrasphaera elongata is Prepadapted to Changing Environmental Conditions in Wastewater Treatment Plants

The activated sludge in wastewater treatment plants (WWTP) designed for enhanced biological phosphorus removal (EBPR) experiences periodically changing nutrient and oxygen availability. Tetrasphaera is the most abundant genus in Danish WWTP and represent up to 20-30% of the activated sludge community based on 16S rRNA amplicon sequencing and quantitative fluorescence in situ hybridization analyses, although it is in low abundance in the influent wastewater. Here we investigate how Tetrasphaera can successfully out-compete most other microorganisms in such highly dynamic ecosystems. To achive this, we analyze the physiological adaptations of the WWTP isolate T. elongata str. LP2 during an aerobic to anoxic shift by label-free quantitative proteomics and NMR-metabolomics. Escherichia coli was used as reference organism as it shares several metabolic capabilities and is regularly introduced to wastewater treatment plants, but without succeeding there. When compared to E. coli, only minor changes in the proteome of T. elongata were observed after the switch to anoxic conditions. This indicates that metabolic pathways for anaerobic energy harvest were already expressed during the aerobic growth. This allows continuous growth of Tetrasphaera immediately after the switch to anoxic conditions. Metabolomics furthermore revealed that the substrates provided were exploited far more efficiently by Tetrasphaera than by E. coli. These results suggest that T. elongata prosper in the dynamic WWTP environment due to adaptation to the changing environmental conditions. Significance of the studyMembers of the genus Tetrasphaera are widely distributed and highly abundant in most well-operating WWTPs with EBPR configuration. However, despite their high abundance in situ, little is known about their physiology and ecological role. Although the importance of Tetrasphaera in engineered wastewater treatment systems is slowly being recognized, additional information is needed to understand the full extent of functions the microorganisms have in many of the essential biological processes in the WWTP. Such information may improve available process models and ultimately lead to better wastewater treatment as well as resource recovery. This study supplies proteomic and metabolomic data on the aerobic/anoxic adaptation of Tetrasphaera and provides a hypothesis on how Tetrasphaera might compete in dynamic engineered systems.

microbiology↗

bioRxiv2018

Resolving the individual contribution of key microbial populations to enhanced biological phosphorus removal with Raman-FISH

Enhanced biological phosphorus removal (EBPR) is a globally important biotechnological process and relies on the massive accumulation of phosphate within special microorganisms. Candidatus Accumulibacter conform to classical physiology model for polyphosphate accumulating organisms and are widely believed to be the most important player for the process in full-scale EBPR systems. However, it was impossible till now to quantify the contribution of specific microbial clades to EBPR. In this study, we have developed a new tool to directly link the identity of microbial cells to the absolute quantification of intracellular poly-P and other polymers under in situ conditions, and applied it to eight full-scale EBPR plants. Besides Ca. Accumulibacter, members of the genus Tetrasphaera were found to be important microbes for P accumulation, and in six plants they were the most important. As these Tetrasphaera cells did not exhibit the classical phenotype of poly-P accumulating microbes, our entire understanding of the microbiology of the EBPR process has to be revised. Furthermore, our new single-cell approach can now also be applied to quantify storage polymer dynamics in individual populations in situ in other ecosystems and might become a valuable tool for many environmental microbiologists.

ecology↗