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富含氨氧化菌的好氧颗粒污泥结构细胞外聚合物中的糖基化淀粉样蛋白。

Glycosylated amyloid-like proteins in the structural extracellular polymers of aerobic granular sludge enriched with ammonium-oxidizing bacteria.

机构信息

Department of Biotechnology, Faculty of Applied Sciences, Delft University of Technology, Delft, The Netherlands.

GENOCOV Research Group, Department of Chemical, Biological and Environmental Engineering, School of Engineering, Universitat Autònoma de Barcelona, Barcelona, Spain.

出版信息

Microbiologyopen. 2018 Dec;7(6):e00616. doi: 10.1002/mbo3.616. Epub 2018 Mar 31.

DOI:10.1002/mbo3.616
PMID:29604180
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6291783/
Abstract

A new type of structural extracellular polymers (EPS) was extracted from aerobic granular sludge dominated by ammonium-oxidizing bacteria. It was analyzed by Raman and FTIR spectroscopy to characterize specific amino acids and protein secondary structure, and by SDS-PAGE with different stains to identify different glycoconjugates. Its intrinsic fluorescence was captured to visualize the location of the extracted EPS in the nitrifying granules, and its hydrogel-forming property was studied by rheometry. The extracted EPS is abundant with cross ß-sheet secondary structure, contains glycosylated proteins/polypeptides, and rich in tryptophan. It forms hydrogel with high mechanical strength. The extraction and discovery of glycosylated proteins and/or amyloids further shows that conventionally used extraction and characterization techniques are not adequate for the study of structural extracellular polymers in biofilms and/or granular sludge. Confirming amyloids secondary structure in such a complex sample is challengeable due to the possibility of amyloids glycosylation and self-assembly. A new definition of extracellular polymers components which includes glycosylated proteins and a better approach to studying them is required to stimulate biofilm research.

摘要

从以氨氧化菌为主的好氧颗粒污泥中提取了一种新型结构细胞外聚合物 (EPS)。通过拉曼和傅里叶变换红外光谱对其进行分析，以表征特定的氨基酸和蛋白质二级结构，并通过 SDS-PAGE 用不同的染色剂来鉴定不同的糖缀合物。通过荧光显微镜观察提取的 EPS 在硝化颗粒中的位置，并通过流变仪研究其水凝胶形成特性。提取的 EPS 富含交叉 β-折叠二级结构，含有糖基化蛋白质/多肽，并且富含色氨酸。它形成具有高机械强度的水凝胶。糖基化蛋白和/或淀粉样蛋白的提取和发现进一步表明，传统的提取和表征技术对于研究生物膜和/或颗粒污泥中的结构细胞外聚合物是不够的。由于淀粉样蛋白的糖基化和自组装的可能性，在如此复杂的样品中确认淀粉样蛋白的二级结构具有挑战性。需要对细胞外聚合物成分进行新的定义，包括糖基化蛋白质，并需要更好的方法来研究它们，以激发生物膜研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7ba7/6291783/96636efea2f6/MBO3-7-e00616-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7ba7/6291783/4d337265a929/MBO3-7-e00616-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7ba7/6291783/07ad6b722d4c/MBO3-7-e00616-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7ba7/6291783/e0b98377cb00/MBO3-7-e00616-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7ba7/6291783/7b653beb4c7b/MBO3-7-e00616-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7ba7/6291783/123533f53974/MBO3-7-e00616-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7ba7/6291783/ca826c9d9cd0/MBO3-7-e00616-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7ba7/6291783/b936847b3363/MBO3-7-e00616-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7ba7/6291783/96636efea2f6/MBO3-7-e00616-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7ba7/6291783/4d337265a929/MBO3-7-e00616-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7ba7/6291783/07ad6b722d4c/MBO3-7-e00616-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7ba7/6291783/e0b98377cb00/MBO3-7-e00616-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7ba7/6291783/7b653beb4c7b/MBO3-7-e00616-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7ba7/6291783/123533f53974/MBO3-7-e00616-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7ba7/6291783/ca826c9d9cd0/MBO3-7-e00616-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7ba7/6291783/b936847b3363/MBO3-7-e00616-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7ba7/6291783/96636efea2f6/MBO3-7-e00616-g008.jpg

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