Investigation of multiple polymers in a denitrifying sulfur conversion-EBPR system: The structural dynamics and...

Polyhydroxyalkanoates (PHAs), polyphosphate (poly-P) and polysulfide or elemental sulfur (poly-S) are the key functionally relevant polymers involved in the recently reported Denitrifying Sulfur conversion-associated Enhanced Biological Phosphorus Removal (DS-EBPR) process. However, little is known about the structural dynamics and storage states of these polymers. In particular, investigating the poly-S generated in this process is quite a superior challenge. This study was thus aimed at simultaneously qualitative-quantitative investigating poly-S and associated poly-P and PHAs through the integrated chemical analysis and Raman micro-spectroscopy coupled with multiple microscopic methods (i.e. optical microscopy, confocal laser scanning microscopy, and differential interference contrast microscopy). The chemical analytical results displayed a stable DS-EBPR phenotype in terms of sulfur conversion, P release/uptake and the dynamics of relevant polymers. The multiple microscopic images and Raman spectrum profiles further clearly demonstrated the existence of the polymers and their dynamic changes under alternating anaerobic-anoxic conditions, consistent with the chemical analytical results. In particular, Raman analysis for the first time unraveled the co-existence of S⁰/S_n^2- species stored either intracellularly or extracellularly; and the dynamic conversions between S⁰/S_n^2- and other sulfur species suggest that there might be a universal pool of bioavailable sulfur. The results reveal the mechanisms underlying the structural dynamics and changes in storage states of the relevant polymers that are functionally relevant to the carbon/phosphorus/sulfur-cycles during different metabolic phases. These mechanisms would otherwise not be obtained only using a traditional chemical analysis-based approach.