Yong-Tao Lv , Xin Zhang , Xiyan Du , Jin Zhang , Lei Wang , Huihui Zhang , Licheng Chen , Lei Wang
{"title":"活性氯对活性污泥反硝化性能的剂量依赖性:细胞外聚合物屏障的构建及酶-底物相互作用机制","authors":"Yong-Tao Lv , Xin Zhang , Xiyan Du , Jin Zhang , Lei Wang , Huihui Zhang , Licheng Chen , Lei Wang","doi":"10.1016/j.jwpe.2025.108858","DOIUrl":null,"url":null,"abstract":"<div><div>When the reclaimed wastewater is used to replenish the lakes and reservoirs, advanced nitrogen removal process is required to prevent the risk of eutrophication. However, the impact of residual active chlorine resulting from NaClO disinfection on activated sludge denitrification systems remains unclear. In this study, the defense mechanisms of extracellular polymeric substances (EPS) and the dynamics of enzyme-substrate interactions were investigated under chlorine stress through batch experiments, combined with spectroscopy and microscale thermophoresis (MST). The results indicated that low concentrations of active chlorine (1–3 mg/L) triggered microbial stress responses, causing a 5.71 %–17.14 % increase in nitrate removal efficiency (NRE) compared to the control group. Notably, the protein components within tightly bound EPS played a key role in resisting chlorine-induced oxidation by forming a hydrophobic barrier. In contrast, high chlorine concentrations (6–12 mg/L) caused oxidative damage to key functional groups of EPS (N<img>H, C<img>O, C<img>O) and compromised the integrity of microbial cell membrane, leading to a 20.00 %–49.86 % decline in NRE. The MST analysis revealed, for the first time, the mechanism through which active chlorine regulates the enzyme-substrate binding affinity. At a low chlorine concentration (1 mg/L), the dissociation constant (K<sub>d</sub>) of nitrate reductase (NAR) and NO<sub>3</sub><sup>−</sup> decreased from 100.85 μM to 23.76 μM, causing a significant rise in the catalytic efficiency of enzyme. In contrast, at high chlorine concentration (12 mg/L), K<sub>d</sub> value increased to 369.46 μM, indicating enzyme deactivation. This study clarifies how chlorine disinfection interferes with activated sludge denitrification. The findings may serve as a theoretical foundation for optimization of advanced nitrogen removal processes in reclaimed water systems.</div></div>","PeriodicalId":17528,"journal":{"name":"Journal of water process engineering","volume":"79 ","pages":"Article 108858"},"PeriodicalIF":6.7000,"publicationDate":"2025-10-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Dose-dependent effects of active chlorine on denitrification performance of activated sludge: Construction of extracellular polymeric substance barrier and enzyme-substrate interaction mechanism\",\"authors\":\"Yong-Tao Lv , Xin Zhang , Xiyan Du , Jin Zhang , Lei Wang , Huihui Zhang , Licheng Chen , Lei Wang\",\"doi\":\"10.1016/j.jwpe.2025.108858\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><div>When the reclaimed wastewater is used to replenish the lakes and reservoirs, advanced nitrogen removal process is required to prevent the risk of eutrophication. However, the impact of residual active chlorine resulting from NaClO disinfection on activated sludge denitrification systems remains unclear. In this study, the defense mechanisms of extracellular polymeric substances (EPS) and the dynamics of enzyme-substrate interactions were investigated under chlorine stress through batch experiments, combined with spectroscopy and microscale thermophoresis (MST). The results indicated that low concentrations of active chlorine (1–3 mg/L) triggered microbial stress responses, causing a 5.71 %–17.14 % increase in nitrate removal efficiency (NRE) compared to the control group. Notably, the protein components within tightly bound EPS played a key role in resisting chlorine-induced oxidation by forming a hydrophobic barrier. In contrast, high chlorine concentrations (6–12 mg/L) caused oxidative damage to key functional groups of EPS (N<img>H, C<img>O, C<img>O) and compromised the integrity of microbial cell membrane, leading to a 20.00 %–49.86 % decline in NRE. The MST analysis revealed, for the first time, the mechanism through which active chlorine regulates the enzyme-substrate binding affinity. At a low chlorine concentration (1 mg/L), the dissociation constant (K<sub>d</sub>) of nitrate reductase (NAR) and NO<sub>3</sub><sup>−</sup> decreased from 100.85 μM to 23.76 μM, causing a significant rise in the catalytic efficiency of enzyme. In contrast, at high chlorine concentration (12 mg/L), K<sub>d</sub> value increased to 369.46 μM, indicating enzyme deactivation. This study clarifies how chlorine disinfection interferes with activated sludge denitrification. The findings may serve as a theoretical foundation for optimization of advanced nitrogen removal processes in reclaimed water systems.</div></div>\",\"PeriodicalId\":17528,\"journal\":{\"name\":\"Journal of water process engineering\",\"volume\":\"79 \",\"pages\":\"Article 108858\"},\"PeriodicalIF\":6.7000,\"publicationDate\":\"2025-10-06\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of water process engineering\",\"FirstCategoryId\":\"5\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S2214714425019312\",\"RegionNum\":2,\"RegionCategory\":\"工程技术\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"ENGINEERING, CHEMICAL\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of water process engineering","FirstCategoryId":"5","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2214714425019312","RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"ENGINEERING, CHEMICAL","Score":null,"Total":0}
Dose-dependent effects of active chlorine on denitrification performance of activated sludge: Construction of extracellular polymeric substance barrier and enzyme-substrate interaction mechanism
When the reclaimed wastewater is used to replenish the lakes and reservoirs, advanced nitrogen removal process is required to prevent the risk of eutrophication. However, the impact of residual active chlorine resulting from NaClO disinfection on activated sludge denitrification systems remains unclear. In this study, the defense mechanisms of extracellular polymeric substances (EPS) and the dynamics of enzyme-substrate interactions were investigated under chlorine stress through batch experiments, combined with spectroscopy and microscale thermophoresis (MST). The results indicated that low concentrations of active chlorine (1–3 mg/L) triggered microbial stress responses, causing a 5.71 %–17.14 % increase in nitrate removal efficiency (NRE) compared to the control group. Notably, the protein components within tightly bound EPS played a key role in resisting chlorine-induced oxidation by forming a hydrophobic barrier. In contrast, high chlorine concentrations (6–12 mg/L) caused oxidative damage to key functional groups of EPS (NH, CO, CO) and compromised the integrity of microbial cell membrane, leading to a 20.00 %–49.86 % decline in NRE. The MST analysis revealed, for the first time, the mechanism through which active chlorine regulates the enzyme-substrate binding affinity. At a low chlorine concentration (1 mg/L), the dissociation constant (Kd) of nitrate reductase (NAR) and NO3− decreased from 100.85 μM to 23.76 μM, causing a significant rise in the catalytic efficiency of enzyme. In contrast, at high chlorine concentration (12 mg/L), Kd value increased to 369.46 μM, indicating enzyme deactivation. This study clarifies how chlorine disinfection interferes with activated sludge denitrification. The findings may serve as a theoretical foundation for optimization of advanced nitrogen removal processes in reclaimed water systems.
期刊介绍:
The Journal of Water Process Engineering aims to publish refereed, high-quality research papers with significant novelty and impact in all areas of the engineering of water and wastewater processing . Papers on advanced and novel treatment processes and technologies are particularly welcome. The Journal considers papers in areas such as nanotechnology and biotechnology applications in water, novel oxidation and separation processes, membrane processes (except those for desalination) , catalytic processes for the removal of water contaminants, sustainable processes, water reuse and recycling, water use and wastewater minimization, integrated/hybrid technology, process modeling of water treatment and novel treatment processes. Submissions on the subject of adsorbents, including standard measurements of adsorption kinetics and equilibrium will only be considered if there is a genuine case for novelty and contribution, for example highly novel, sustainable adsorbents and their use: papers on activated carbon-type materials derived from natural matter, or surfactant-modified clays and related minerals, would not fulfil this criterion. The Journal particularly welcomes contributions involving environmentally, economically and socially sustainable technology for water treatment, including those which are energy-efficient, with minimal or no chemical consumption, and capable of water recycling and reuse that minimizes the direct disposal of wastewater to the aquatic environment. Papers that describe novel ideas for solving issues related to water quality and availability are also welcome, as are those that show the transfer of techniques from other disciplines. The Journal will consider papers dealing with processes for various water matrices including drinking water (except desalination), domestic, urban and industrial wastewaters, in addition to their residues. It is expected that the journal will be of particular relevance to chemical and process engineers working in the field. The Journal welcomes Full Text papers, Short Communications, State-of-the-Art Reviews and Letters to Editors and Case Studies