Nitrogen removal characteristics and bioenhancement effects of aerobic denitrifying bacteria Burkholderia sp. ZH8
LI Yan-li, YANG Lei, ZHANG Zhi-hao, LI Yu-cai, MENG Hong-yan, REN Yong-xiang
Shaanxi Key Laboratory of Environmental Engineering, Key Laboratory of Northwest Water Resource, Environment and Ecology, Ministry of Education, Xi'an University of Architecture and Technology, Xi'an 710055, China
Abstract：Aiming at the problems of complicated process and low denitrification efficiency of traditional biological denitrification treatment of nitrogen-containing wastewater, an aerobic denitrification strain ZH8 was screened from a stable biological filter in the laboratory, and this strain had strong heterotrophic nitrification and aerobic denitrification ability. Strain ZH8was identified as Burkholderia sp. by morphological, physiological and biochemical characteristics and 16S rDNA sequence analysis. The single factor experiments showed that the optimal culture conditions of aerobic denitrification strain ZH8were as follows: carbon source was sodium succinate, C/N was 15, temperature was 30℃, rotational speed was 160r/min and pH was 7~8. In addition, strain ZH8 could not only grow with NH4+-N, NO3--N or NO2--N as a single nitrogen source, but also achieve simultaneous nitrification and denitrification under mixed nitrogen source conditions. According to the whole genome analysis, heterotrophic nitrification of strain ZH8 was ammonium assimilation through glutamate dehydrogenase pathway, and aerobic denitrification pathway was NO3--N→NO2--N→NO→N2O→N2. Carbon metabolism pathway included tricarboxylic acid cycle, glyoxylic acid cycle, glycolysis and pentose phosphate. In the biofilm reactor cultured with pure bacterium ZH8, the reaction could be completed after 8 to 15 days, and the system could maintain efficient and stable nitrogen removal performance, and the removal rates of NH4+-N, NO3--N and TN reached more than 90%. The strain provides a new strain resource for wastewater bio-treatment, which can effectively treat nitrogen-containing wastewater and has practical application value.
[1] 卢瑞朋,徐文江,李安峰,等.强化反硝化除磷的新型多级缺氧-好氧工艺[J]. 中国环境科学, 2022,42(4):8. Lu R P, Xu W J, Li A F, et al. Novel multi-stage anoxic-aerobic process for enhanced denitrification and phosphorus removal [J]. China Environmental Science, 2022,42(4):1706-1713. [2] 郭超,尹辉,范奎,等.一株好氧反硝化菌的分离鉴定及脱氮特性研究[J]. 安全与环境工程, 2020,27(4):41-47. Guo C, Yin H, Fan K, et al. Isolation and Identification of an Aerobic Denitrifying Bacteria and Its Denitrification Characteristics [J]. Safety and Environmental Engineering, 2020,27(4):41-47. [3] Ren J, Cheng X, Ma H, et al. Characteristics of a novel heterotrophic nitrification and aerobic denitrification bacterium and its bioaugmentation performance in a membrane bioreactor [J]. Bioresource Technology, 2021,342:125908. [4] 冯亮,袁春燕,杨超,等.好氧反硝化生物脱氮技术的研究进展[J]. 微生物学通报, 2020,47(10):3342-3354. Feng L, Yuan C Y, Yang C, et al. Research progress of aerobic denitrification biological denitrification technology [J]. Microbiology Chinese Bulletin, 2020,47(10):3342-3354. [5] 郭焱,何雯,吴丹,等.好氧反硝化菌Pseudomonas sp. J-5的分离及脱氮性能研究[J]. 绿色科技, 2023,25(14):224-229. Guo Y, He W, Wu D, et al. Isolation and denitrification performance of aerobic denitrifying bacterium Pseudomonas sp.J-5[J]. Green Technology, 2023,25(14):224-229. [6] Xie F, Thiri M, Wang H. Simultaneous heterotrophic nitrification and aerobic denitrification by a novel isolated Pseudomonas mendocina X49[J]. Bioresource Technology, 2021,319:124198. [7] 赵洋,孙慧明,林浩澎,等.一株安全高效的好氧反硝化菌Pseudomonas stutzeri DZ11的生物安全性及脱氮性能研究[J]. 生物技术通报, 2022,38(10):226. Zhao Y, Sun H M, Lin H P, et al. Study on Biosafety and Denitrification performance of Pseudomonas stutzeri DZ11, a safe and efficient aerobic denitrification bacterium [J]. Bulletin of Biotechnology, 2022,38(10):226. [8] 东秀珠,蔡妙英.常见细菌系统鉴定手册[M]. 北京:科学出版社, 2001. Dong X Z, Cai M Y. Handbook of Systematic Identification of Common Bacteria [M]. Beijing: Science Press, 2001. [9] Chen J, Xu J, Zhang S, et al. Nitrogen removal characteristics of a novel heterotrophic nitrification and aerobic denitrification bacteria, Alcaligenes faecalis strain WT14[J]. Journal of environmental management, 2021,282:111961. [10] 陈均利,张苗苗,张树楠,等. Alcaligenes faecalis WT14的异养硝化-好氧反硝化特性及对高氨废水处理潜力[J]. 环境工程, 2021,39(2): 27-32,40. Chen J L, Zhang M M, Zhang S N, et al. Heterotrophic nitrification and aerobic denitrification characteristics of Alcaligenes faecalis WT14 and its potential for treatment of high ammonia wastewater [J]. Environmental Engineering, 2021,39(2):27-32,40. [11] 沈辉,万夕和,蒋葛,等.1株滩涂沉积物反硝化细菌的鉴定及其性能研究[J]. 微生物学杂志, 2016,36(2):50-55. Shen H, Wan X H, Jiang G, et al. Identification and performance of denitrifying bacteria from mudflat sediments [J]. Chinese Journal of Microbiology, 2016,36(2):50-55. [12] 王永刚,王旭,张俊娥,等.好氧反硝化细菌研究及应用进展[J]. 工业水处理, 2017,37(2):12-17. Wang Y G, Wang X, Zhang J E, et al. Research progress and application of aerobic denitrifying bacteria [J]. Industrial Water Treatment, 2017,37(2):12-17. [13] Zhao B, An Q, He Y L, et al. N2O and N2production during heterotrophic nitrification by Alcaligenes faecalis strain NR [J]. Bioresource Technology, 2012,116:379-385. [14] Zhu L, Ding W, Feng L, et al. Characteristics of an aerobic denitrifier that utilizes ammonium and nitrate simultaneously under the oligotrophic niche [J]. Environmental Science and Pollution Research, 2012,19:3185-3191. [15] Wehrfritz J M, Reilly A, Spiro S, et al. Purification of hydroxylamine oxidase from Thiosphaera pantotropha: Identification of electron acceptors that couple heterotrophic nitrification to aerobic denitrification [J]. FEBS letters, 1993,335(2):246-250. [16] Zhang M, Anzhang, et al. Nitrogen removal characteristics of a versatile heterotrophic nitrifying-aerobic denitrifying bacterium, Pseudomonas bauzanensis DN13-1, isolated from deep-sea sediment [J]. Bioresource technology, 2019,305:122626-122626. [17] 胡家玮,李军,卞伟,等.城市污水连续流A/O系统富氧条件下脱氮特征[J]. 化工学报, 2014,65(10):4071-4077. Hu J W, Li J, Bian W, et al. Characteristics of nitrogen removal in continuous flow A/O system [J]. Acta Chemologica Sinica, 2014, 65(10):4071-4077. [18] Ge Q, Yue X, Wang G. Simultaneous heterotrophic nitrification and aerobic denitrification at high initial phenol concentration by isolated bacterium Diaphorobacter sp. PD-7[J]. Chinese Journal of Chemical Engineering, 2015,23(5):835-841. [19] 姜磊,徐成斌,马溪平,等.1株好氧反硝化菌的分离鉴定和反硝化特性研究[J]. 环境科学与技术, 2013,36(3):12-15. Jiang L, Xu C B, Ma X P, et al. Isolation, identification and denitrification characteristics of a strain of aerobic denitrifying bacteria [J]. Environmental Science and Technology, 2013,36(3):12-15. [20] 刘小英,冯晟,班宜辉,等.一株异养硝化-好氧反硝化细菌的分离鉴定及其脱氮性能研究[J]. 生态环境学报, 2016,25(12):1983-1990. Liu X Y, Feng S, Ban Y H, et al. Isolation, Identification and denitrification performance of a heterotrophic nitrifying aerobic denitrifying bacteria [J]. Journal of Ecology and Environment, 2016,25(12):1983-1990. [21] Chen Q, Ni J. Heterotrophic nitrification-aerobic denitrification by novel isolated bacteria [J]. Journal of Industrial Microbiology and Biotechnology, 2011,38(9):1305-1310. [22] Li C, Yang J, Wang X, et al. Removal of nitrogen by heterotrophic nitrification-aerobic denitrification of a phosphate accumulating bacterium Pseudomonas stutzeri YG-24[J]. Bioresource Technology, 2015,182:18-25. [23] Huang F, Pan L, He Z, et al. Culturable heterotrophic nitrification- aerobic denitrification bacterial consortia with cooperative interactions for removing ammonia and nitrite nitrogen in mariculture effluents [J]. Aquaculture, 2020:735211. [24] Sun Z, Lv Y, Liu Y, et al. Removal of nitrogen by heterotrophic nitrification-aerobic denitrification of a novel metal resistant bacterium Cupriavidus sp. S1[J]. Bioresour Technology, 2016:142-150. [25] Chen Q, Ni J. Ammonium removal by Agrobacterium sp. LAD9 capable of heterotrophic nitrification-aerobic denitrification [J]. Journal of Bioscience and Bioengineering, 2012,113(5). [26] Yao S, Ni J, Chen Q, et al. Enrichment and characterization of a bacteria consortium capable of heterotrophic nitrification and aerobic denitrification at low temperature [J]. Bioresource technology, 2013, 127:151-157. [27] Wang T, Liu Z X, Wu M J, et al. Screening and Characterization of a Bacterium Capable of Simultaneous Heterotrophic Nitrification and Aerobic Denitrification at High Concentrations of Ammonia-Nitrogen [J]. Applied Mechanics and Materials, 2014,665:487-490. [28] 杨垒,陈宁,任勇翔,等.异养硝化细菌Acinetobacter junii NP1的同步脱氮除磷特性及动力学分析[J]. 环境科学, 2019,40(8):3713-3721. Yang L, Chen N, Ren Y X, et al. Simultaneous nitrogen and phosphorus removal by heterotrophic nitrifying bacteria Acinetobacter junii NP1[J]. Environmental Science, 2019,40(8):3713-3721. [29] 梁贤,任勇翔,杨垒,等.异养硝化-好氧反硝化菌YL的脱氮特性[J]. 环境科学, 2015,(5):1749-1756. Liang X, Ren Y X, Yang L, et al. Nitrogen removal characteristics of heterotrophic nitrifying and aerobic denitrifying bacteria YL [J]. Environmental Science, 2015,36(5):1749-1756. [30] 何腾霞,李振轮,徐义.耐冷亚硝酸盐型反硝化细菌Pseudomonas putida Y-12脱氮特性[J]. 环境科学学报, 2015,35(10):3071-3077. He T X, Li Z L, Xu Y. Nitrogen removal characteristics of cold nitrite-resistant denitrifying bacterium Pseudomonas putida Y-12[J]. Journal of Environmental Sciences, 2015,35(10):3071-3077. [31] Jin P, Chen Y, Yao R, et al. New insight into the nitrogen metabolism of simultaneous heterotrophic nitrification-aerobic denitrification bacterium in mRNA expression [J]. Journal of hazardous materials, 2019,371:295-303. [32] Pal R R, Khardenavis A A, Purohit H J. Identification and monitoring of nitrification and denitrification genes in Klebsiella pneumoniae EGD-HP19-C for its ability to perform heterotrophic nitrification and aerobic denitrification [J]. Functional & integrative genomics, 2015, 15(1):63-76. [33] Bai Y, Su J, Ali A, et al. Insights into the mechanism of Mn (II)-based autotrophic denitrification: Performance, genomic, and metabonomics [J]. Science of The Total Environment, 2022,810:151185. [34] Hasan M T, Sun A, Khatiwada B, et al. Comparative proteomics investigation of central carbon metabolism in Euglena gracilis grown under predominantly phototrophic, mixotrophic and heterotrophic cultivations [J]. Algal Research, 2019,43:101638. [35] Fu X, Hou R, Yang P, et al. Application of external carbon source in heterotrophic denitrification of domestic sewage: A review [J]. Science of The Total Environment, 2022,817:153061. [36] Zhang Q, Chen X, Zhang Z, et al. Performance and microbial ecology of a novel moving bed biofilm reactor process inoculated with heterotrophic nitrification-aerobic denitrification bacteria for high ammonia nitrogen wastewater treatment [J]. Bioresource technology, 2020,315:123813. [37] He X, Sun Q, Xu T, et al. Removal of nitrogen by heterotrophic nitrification-aerobic denitrification of a novel halotolerant bacterium Pseudomonas mendocina TJPU04[J]. Bioprocess and biosystems engineering, 2019,42(5):853-866. [38] Duan J, Fang H, Su B, et al. Characterization of a halophilic heterotrophic nitrification-aerobic denitrification bacterium and its application on treatment of saline wastewater [J]. Bioresource technology, 2015,179:421-428.
