Characterization of extracellular polymer substances and acyl-homoserine lactones of sulfur-based autotrophic denitrification microbial community under high nitrate loading
XU Wei-yi1, HUANG Kai-wen1, LIU Chong2, SHEN Fei3, WANG Shuo1,4, LI Ji1,4
1. Jiangsu Key Laboratory of Anaerobic Biotechnology, School of Environment & Ecology, Jiangnan University, Wuxi 214122, China; 2. 101 Institute of the Ministry of Civil Affairs, Beijing 100070, China; 3. Laboratory of Instrumental Analysis, Jiangsu Wuxi Environmental Monitoring Center, Wuxi 214121, China; 4. Jiangsu College of Water Treatment Technology and Material Collaborative Innovation Center, Suzhou 215009, China
Abstract：Sulfur-based autotrophic denitrification (SADN) was influenced by hydraulic retention time (HRT) and influent NO3--N concentration. By progressively shortening the HRT and increasing the influent NO3--N concentration, the nitrogen removal limit of the process was explored. Simultaneously, the changes in extracellular polymeric substances (EPS) and signal molecules acyl-homoserine lactones (AHLs) were analyzed, along with the evolution of microbial community structure in this process. During the transition from an HRT of 4.1h to 1.0h, SADN bacteria rapidly adapted to the environment. When the HRT was 1.0h and the influent NO3--N concentration was 40mg/L, the system achieved a TN removal rate of 99.74%, with a denitrification load as high as 958.53mg/(L·d). In this process, the increase in EPS concentration was mainly driven by TB-PN and TB-PS, while the increase in AHLs concentration was primarily led by C4-HSL. In comparison to shortening the HRT, raising the influent NO3--N concentration significantly increased the C4-HSL concentration, leading to a gradual trend towards an unstable state in the sludge structure. C6-HSL, the predominant AHL in the system, exhibited substantial growth in stages G4 and G7, reflecting a self-regulation by the microorganisms following the weakening of the system's denitrification capability. Analysis of the microbial community composition indicated that system operation and nitrogen load variations drove the domestication selection of microbial communities. In the reduced HRT group, norank_f_ Pleomorphomonadaceae of Proteobacteria played a primary role as the dominant bacterial genus, providing substrates for denitrification bacteria to sustain system stability. Following the increase in NO3--N concentration, UKL 13-1, Simplicispira, Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium, and Thermomonas gradually evolved into dominant bacterial genera, all crucial microorganisms with denitrification capabilities.
许炜怡, 黄凯文, 刘崇, 沈斐, 王硕, 李激. 高氮负荷下硫自养反硝化菌群的胞外聚合物和信号分子特征[J]. 中国环境科学, 2024, 44(8): 4314-4325.
XU Wei-yi, HUANG Kai-wen, LIU Chong, SHEN Fei, WANG Shuo, LI Ji. Characterization of extracellular polymer substances and acyl-homoserine lactones of sulfur-based autotrophic denitrification microbial community under high nitrate loading. CHINA ENVIRONMENTAL SCIENCECE, 2024, 44(8): 4314-4325.
[1] Qu J, Fan M. The current state of water quality and technology development for water pollution control in China [J]. Critical Reviews in Environmental Science and Technology, 2010,40(6):519-560. [2] Cheng S S, Chen W C. Organic carbon supplement influencing performance of biological nitritification in a fluidized bed reactor [J]. Water Science and Technology, 1994,30(11):131-142. [3] Bisogni J J J, Driscoll Jr C T. Denitrification using thiosulfate and sulfide [J]. Journal of the Environmental Engineering division, 1977, 103(4):593-604. [4] Shao M F, Zhang T, Fang H H. Sulfur-driven autotrophic denitrification: diversity, biochemistry, and engineering applications [J]. Applied Microbiology and Biotechnology, 2010,88(5):1027-1042. [5] Kostrytsia A, Papirio S, Frunzo L, et al. Elemental sulfur-based autotrophic denitrification and denitritation: microbially catalyzed sulfur hydrolysis and nitrogen conversions [J]. Journal of Environmental Management, 2018,211:313-322. [6] Sahinkaya E, Kilic A. Heterotrophic and elemental-sulfur-based autotrophic denitrification processes for simultaneous nitrate and Cr(VI) reduction [J]. Water Research, 2014,50:278-286. [7] Cui Y X, Biswal B K, van Loosdrecht M C M, et al. Long term performance and dynamics of microbial biofilm communities performing sulfur-oxidizing autotrophic denitrification in a moving- bed biofilm reactor [J]. Water Research, 2019,166:115038-115051. [8] Tolhurst T J. Weak diurnal changes in the biochemical properties and benthic macrofauna of urbanised mangrove forests and mudflats [J]. Hydrobiologia, 2009,636(1):101-117. [9] Boleij M, Pabst M, Neu T R, et al. Identification of glycoproteins isolated from extracellular polymeric substances of full-scale anammox granular sludge [J]. Environmental Science and Technology, 2018,52:13127-13135. [10] Zhang J, Miao Y, Zhang Q, et al. Mechanism of stable sewage nitrogen removal in a partial nitrification-anammox biofilm system at low temperatures: microbial community and EPS analysis [J]. Bioresource Technology, 2020,297:122459-122465. [11] Zhang Z, Wang L, Ji Y, et al. Understanding the N-acylated homoserine lactones(AHLs)-based quorum sensing for the stability of aerobic granular sludge in the aspect of substrate hydrolysis enhancement [J]. Science of the Total Environment, 2023,858: 159581-159596. [12] Chen F, Li X, Gu C, et al. Selectivity control of nitrite and nitrate with the reaction of S0and achieved nitrite accumulation in the sulfur autotrophic denitrification process [J]. Bioresource Technology, 2018, 266:211-219. [13] Song C K, Wang Y Y, Han H C, et al. Effect of decreasing temperature on the performance and extracellular polymer substance of anaerobic ammonia oxidation sludge [J]. China Environmental Sciencece, 2016, 36:2006-2013. [14] Hou X, Liu S, Zhang Z. Role of extracellular polymeric substance in determining the high aggregation ability of anammox sludge [J]. Water Research, 2015,75:51-62. [15] Zhang S, Zeng X, Ren M, et al. Novel metabolic and physiological functions of branched chain amino acids: a review [J]. Journal of Animal Science and Biotechnology, 2017,8(1):10-22. [16] Tang X, Guo Y, Wu S, et al. Metabolomics uncovers the regulatory pathway of acyl-homoserine lactones based quorum sensing in anammox consortia [J]. Environmental Science & Technology, 2018, 52(4):2206-2216. [17] Zhang B, Lens P N L, Shi W, et al. The attachment potential and N-acyl-homoserine lactone-based quorum sensing in aerobic granular sludge and algal-bacterial granular sludge [J]. Applied Microbiology and Biotechnology, 2018,102(12):5343-5353. [18] Kugimiya A, Kohara K. Amino acid sensing using an ion-sensitive field-effect transistor [J]. Chemical Engineering Journal, 2012,6: 397-400. [19] 徐金梁.氨基酸对厌氧氨氧化系统的影响及机制研究[D]. 2022. Xu J L. Study on the effect and mechanism of amino acids on anammox system [D]. 2022. [20] Husna A U, Wang N, Cobbold S A, et al. Methionine biosynthesis and transport are functionally redundant for the growth and virulence of salmonella typhimurium [J]. Journal of Biological Chemistry, 2018, 293(24):9506-9519. [21] Wen C, Paul W, Leenheer J A, et al. Fluorescence excitation-emission matrix regional integration to quantify spectra for dissolved organic matter [J]. Environmental science & technology, 2003,37(24):5701- 5710. [22] Parthasarathy A, Cross P J, Dobson R C J, et al. A three-ring circus: metabolism of the three proteogenic aromatic amino acids and their role in the health of plants and animals [J]. Frontiers in Molecular Biosciences, 2018,5:29-59. [23] Zhang T, Cao J, Zhu Q, et al. Revealing the characteristics and formation mechanisms of partial denitrification granular sludge for efficient nitrite accumulation driven by glycerol [J]. Chemical Engineering Journal, 2022,428:131195-131204. [24] Chen X, Hu X, Lu Q, et al. Study on the differences in sludge toxicity and microbial community structure caused by catechol, resorcinol and hydroquinone with metagenomic analysis [J]. Journal of Environmental Management, 2022,302:114027-114037. [25] Wei L, Li J, Xue M, et al. Adsorption behaviors of Cu2+, Zn2+ and Cd2+ onto proteins, humic acid, and polysaccharides extracted from sludge EPS: Sorption properties and mechanisms [J]. Bioresource Technology, 2019,291:121868-121877. [26] Wang W, Xie H, Wang H, et al. Organic compounds evolution and sludge properties variation along partial nitritation and subsequent anammox processes treating reject water [J]. Water Research, 2020,184:116197-116209. [27] Badireddy A R, Chellam S, Gassman P L, et al. Role of extracellular polymeric substances in bioflocculation of activated sludge microorganisms under glucose-controlled conditions [J]. Water Research, 2010,44(15):4505-4516. [28] Yang F, Qu J, Huang X, et al. Phosphorus deficiency leads to the loosening of activated sludge: The role of exopolysaccharides in aggregation [J]. Chemosphere, 2022,290:133385-133395. [29] Wen H Q, Ren H Y, Xie G J, et al. Synthesized effects of proteomic and extracellular polymeric substance (EPS) revealing the enhanced hydrogen production by formed biofilm of photo-fermentative bacteria [J]. Environment International, 2020,139:105683-105698. [30] Sang W, Li X, Feng Y, et al. Improvement of the sludge flocculation dewatering efficient by electromagnetic wave loading: research based on removal of bound water [J]. Environmental Science and Pollution Research, 2020,27(3):3413-3427. [31] Zhao H Z, Sun J J, Song J, et al. Direct electron transfer and conformational change of glucose oxidase on carbon nanotube-based electrodes [J]. Carbon, 2010,48(5):1508-1514. [32] Gao H, Zhao R, Wu Z, et al. New insights into exogenous N-acyl- homoserine lactone manipulation in biological nitrogen removal system against ZnO nanoparticle shock [J]. Bioresource Technology, 2022:128567-128576. [33] Ahumedo Monterrosa M, Galindo J F, Vergara Lorduy J, et al. The role of lasR active site amino acids in the interaction with the acyl homoserine lactones (AHLs) analogues: A computational study [J]. Journal of Molecular Graphics and Modelling, 2019,86:113-124. [34] Tan C H, Koh K S, Xie C, et al. The role of quorum sensing signalling in EPS production and the assembly of a sludge community into aerobic granules [J]. Isme Journal, 2014,8(6):1186-1197. [35] Zhang Z, Cao R, Jin L, et al. The regulation of N-acyl-homoserine lactones (AHLs)-based quorum sensing on EPS secretion via ATP synthetic for the stability of aerobic granular sludge [J]. Science of the Total Environment, 2019,673:83-91. [36] Wang J, Liu Q, Li X, et al. In-situ monitoring AHL-mediated quorum-sensing regulation of the initial phase of wastewater biofilm formation [J]. Environment International, 2020,135:105326-105335. [37] Valle J, Toledo Arana A, Berasain C, et al. SarA and not sigmaB is essential for biofilm development by Staphylococcus aureus [J]. Molecular Microbiology, 2003,48(4):1075-1087. [38] Lu Y, Zhang X, Feng L, et al. Optimization of continuous-flow solid-phase denitrification via coupling carriers in enhancing simultaneous removal of nitrogen and organics for agricultural runoff purification [J]. Biodegradation, 2017,28(4):275-285. [39] Chu L, Wang J. Denitrification performance and biofilm characteristics using biodegradable polymers PCL as carriers and carbon source [J]. Chemosphere, 2013,91(9):1310-1316. [40] Ucar D, Yilmaz T, Di Capua F, et al. Comparison of biogenic and chemical sulfur as electron donors for autotrophic denitrification in sulfur-fed membrane bioreactor (SMBR) [J]. Bioresource Technology, 2020,299:122574-122581.
