Elucidating the complex hydrolysis and conversion network of xanthan-like extracellular heteropolysaccharides in waste activated sludge fermentation

The hydrolysis of structural extracellular polymeric substances (St-EPS) is considered a major limiting step in the anaerobic fermentation of waste activated sludge (WAS). However, the degradation of heteropolysaccharides, characterized by complex monomers of uronic acids and neutral saccharides in...

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Main Authors: Chen-Yuan Zhou, Kun Dai, Yi-Peng Lin, Xing-Chen Huang, Yan-Lin Hu, Xuan-Xin Chen, Xiao-Fei Yang, Qi-Yuan Sun, Yong Zhang, Mark C.M. van Loosdrecht, Raymond Jianxiong Zeng, Fang Zhang
Format: Article
Language:English
Published: Elsevier 2025-05-01
Series:Water Research X
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Online Access:http://www.sciencedirect.com/science/article/pii/S2589914725000039
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author Chen-Yuan Zhou
Kun Dai
Yi-Peng Lin
Xing-Chen Huang
Yan-Lin Hu
Xuan-Xin Chen
Xiao-Fei Yang
Qi-Yuan Sun
Yong Zhang
Mark C.M. van Loosdrecht
Raymond Jianxiong Zeng
Fang Zhang
author_facet Chen-Yuan Zhou
Kun Dai
Yi-Peng Lin
Xing-Chen Huang
Yan-Lin Hu
Xuan-Xin Chen
Xiao-Fei Yang
Qi-Yuan Sun
Yong Zhang
Mark C.M. van Loosdrecht
Raymond Jianxiong Zeng
Fang Zhang
author_sort Chen-Yuan Zhou
collection DOAJ
description The hydrolysis of structural extracellular polymeric substances (St-EPS) is considered a major limiting step in the anaerobic fermentation of waste activated sludge (WAS). However, the degradation of heteropolysaccharides, characterized by complex monomers of uronic acids and neutral saccharides in St-EPS, has rarely been reported. In this study, microbial-produced xanthan-like heteropolysaccharides, characterized by a blue filamentary film, were identified. The xanthan-producing bacteria comprised ∼7.2% of total genera present in WAS. An xanthan-degrading consortium (XDC) was enriched in an anaerobic batch reactor. This consortium could degrade Xanthan for over 90% and disrupt the gel structure of xanthan while promoting methane production from WAS by 29%. The xanthan degradation network consisting of extracellular enzymes and bacteria was elucidated by combining high-throughput sequencing, metagenomic, and metaproteomic analyses. Five enzymes were identified as responsible for hydrolyzing xanthan to monomers, including xanthan lyase, β-d-glucosidase, β-d-glucanase, α-d-mannosidase, and unsaturated glucuronyl hydrolase. Seven genera, including Paenibacillus (0.2%) and Clostridium (3.1%), were identified as key bacteria excreting one to five of the aforementioned enzymes. This study thus provides insights into the complex conversions in anaerobic digestion of WAS and gives a foundation for future optimization of this process.
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publishDate 2025-05-01
publisher Elsevier
record_format Article
series Water Research X
spelling doaj-art-e06131af3d884529977fc269cb4563532025-01-16T04:29:11ZengElsevierWater Research X2589-91472025-05-0127100303Elucidating the complex hydrolysis and conversion network of xanthan-like extracellular heteropolysaccharides in waste activated sludge fermentationChen-Yuan Zhou0Kun Dai1Yi-Peng Lin2Xing-Chen Huang3Yan-Lin Hu4Xuan-Xin Chen5Xiao-Fei Yang6Qi-Yuan Sun7Yong Zhang8Mark C.M. van Loosdrecht9Raymond Jianxiong Zeng10Fang Zhang11Center of Wastewater Resource Recovery, College of Resources and Environment, Fujian Agriculture and Forestry University, Fuzhou, Fujian 350002, ChinaCenter of Wastewater Resource Recovery, College of Resources and Environment, Fujian Agriculture and Forestry University, Fuzhou, Fujian 350002, ChinaCenter of Wastewater Resource Recovery, College of Resources and Environment, Fujian Agriculture and Forestry University, Fuzhou, Fujian 350002, ChinaCenter of Wastewater Resource Recovery, College of Resources and Environment, Fujian Agriculture and Forestry University, Fuzhou, Fujian 350002, ChinaCenter of Wastewater Resource Recovery, College of Resources and Environment, Fujian Agriculture and Forestry University, Fuzhou, Fujian 350002, ChinaCenter of Wastewater Resource Recovery, College of Resources and Environment, Fujian Agriculture and Forestry University, Fuzhou, Fujian 350002, ChinaCenter of Wastewater Resource Recovery, College of Resources and Environment, Fujian Agriculture and Forestry University, Fuzhou, Fujian 350002, ChinaCollege of Environmental Science and Engineering, Fujian Normal University, Fuzhou, Fujian 350007, ChinaCollege of Environmental Science and Engineering, Fujian Normal University, Fuzhou, Fujian 350007, ChinaDepartment of Biotechnology, Delft University of Technology, Julianalaan 67, Delft 2628 BC, the NetherlandsCenter of Wastewater Resource Recovery, College of Resources and Environment, Fujian Agriculture and Forestry University, Fuzhou, Fujian 350002, ChinaCenter of Wastewater Resource Recovery, College of Resources and Environment, Fujian Agriculture and Forestry University, Fuzhou, Fujian 350002, China; Corresponding author.The hydrolysis of structural extracellular polymeric substances (St-EPS) is considered a major limiting step in the anaerobic fermentation of waste activated sludge (WAS). However, the degradation of heteropolysaccharides, characterized by complex monomers of uronic acids and neutral saccharides in St-EPS, has rarely been reported. In this study, microbial-produced xanthan-like heteropolysaccharides, characterized by a blue filamentary film, were identified. The xanthan-producing bacteria comprised ∼7.2% of total genera present in WAS. An xanthan-degrading consortium (XDC) was enriched in an anaerobic batch reactor. This consortium could degrade Xanthan for over 90% and disrupt the gel structure of xanthan while promoting methane production from WAS by 29%. The xanthan degradation network consisting of extracellular enzymes and bacteria was elucidated by combining high-throughput sequencing, metagenomic, and metaproteomic analyses. Five enzymes were identified as responsible for hydrolyzing xanthan to monomers, including xanthan lyase, β-d-glucosidase, β-d-glucanase, α-d-mannosidase, and unsaturated glucuronyl hydrolase. Seven genera, including Paenibacillus (0.2%) and Clostridium (3.1%), were identified as key bacteria excreting one to five of the aforementioned enzymes. This study thus provides insights into the complex conversions in anaerobic digestion of WAS and gives a foundation for future optimization of this process.http://www.sciencedirect.com/science/article/pii/S2589914725000039Xanthan-degrading consortiumHeteropolysaccharidesDegradation networkXanthan lyaseMethane production
spellingShingle Chen-Yuan Zhou
Kun Dai
Yi-Peng Lin
Xing-Chen Huang
Yan-Lin Hu
Xuan-Xin Chen
Xiao-Fei Yang
Qi-Yuan Sun
Yong Zhang
Mark C.M. van Loosdrecht
Raymond Jianxiong Zeng
Fang Zhang
Elucidating the complex hydrolysis and conversion network of xanthan-like extracellular heteropolysaccharides in waste activated sludge fermentation
Water Research X
Xanthan-degrading consortium
Heteropolysaccharides
Degradation network
Xanthan lyase
Methane production
title Elucidating the complex hydrolysis and conversion network of xanthan-like extracellular heteropolysaccharides in waste activated sludge fermentation
title_full Elucidating the complex hydrolysis and conversion network of xanthan-like extracellular heteropolysaccharides in waste activated sludge fermentation
title_fullStr Elucidating the complex hydrolysis and conversion network of xanthan-like extracellular heteropolysaccharides in waste activated sludge fermentation
title_full_unstemmed Elucidating the complex hydrolysis and conversion network of xanthan-like extracellular heteropolysaccharides in waste activated sludge fermentation
title_short Elucidating the complex hydrolysis and conversion network of xanthan-like extracellular heteropolysaccharides in waste activated sludge fermentation
title_sort elucidating the complex hydrolysis and conversion network of xanthan like extracellular heteropolysaccharides in waste activated sludge fermentation
topic Xanthan-degrading consortium
Heteropolysaccharides
Degradation network
Xanthan lyase
Methane production
url http://www.sciencedirect.com/science/article/pii/S2589914725000039
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