Journal
CHEMOSPHERE
Volume 259, Issue -, Pages -Publisher
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.chemosphere.2020.127539
Keywords
Extracellular polymeric substances; Anammox granule; Extraction method; Protein secondary structure; Circular dichroism
Categories
Funding
- National Natural Science Foundation of China [51778446, 51522809]
- Foundation of the State Key Laboratory of Pollution Control and Resource Reuse (Tongji University, China) [PCRRY 0400231010]
- National Engineering Laboratory for Advanced Municipal Wastewater Treatment and Reuse Technology
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Extracellular proteins are crucial for the formation and structural maintenance of anammox granules. However, the influence of extracellular polymeric substances (EPS) extraction methods on the secondary structure of extracellular proteins has not been studied fully. In this work, ten EPS extraction methods were applied to anammox granules and compared systematically. The EPS extraction methods were then applied to bovine serum albumin (BSA), a representative of extracellular proteins, to investigate their influence on protein secondary structure by circular dichroism (CD) spectroscopy. The ten EPS extraction methods yielded EPS concentration varying from 5.2 +/- 0.1 to 84.8 +/- 2.0 mg/g volatile suspended solid (VSS). Tween-20 treatment resulted in the most similar CD spectrum and protein secondary structure to the control. Treatment 0.5% Tween-20 for 4 h was found to be the best condition to preserve protein secondary structure while achieving a high EPS yield (44.4 +/- 1.4 mg/g VSS) and limited cell lysis for anammox granules. This study provides an optimized protocol to extract granular EPS when the role of the secondary structure of extracellular proteins in regulating aggregation, adsorption and biofilm formation of microbes is to be elucidated. (C) 2020 Elsevier Ltd. All rights reserved.
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