Screening and application of highly-efficient degrading strain for N-methyl-2-pyrrolidone (NMP)
LI Ning-ning1,2,3, Lü Hai-hui3, YU De-shui3, LI Yu-tong3, WANG Ye3, KANG Jian-ming1,2, XUE Yu-xin3, JIN Yan1, HAN Xu-shen1, YU Jian-guo2,3
1. National Engineering Research Center for Integrated Utilization of Salt Lake Resources, East China University of Science and Technology, Shanghai 200237, China; 2. Joint International Laboratory for Potassium and Lithium Strategic Resources, East China University of Science and Technology, Shanghai 200237, China; 3. School of Chemical Engineering, East China University of Science and Technology, Shanghai 200237, China
Abstract：N-methyl-2-pyrrolidone (NMP) was selected as the sole carbon and nitrogen source, and a strain NCSL-HH10 that could efficiently degrade NMP was isolated from the cleaning wastewater of lithium-ion battery cathode slurry mixer. 16S rDNA sequencing and phylogenetic affiliation analysis showed that this strain belonged to Burkholderia contaminans. The results showed that 100% NMP removal and 94.3% TOC removal could be obtained in 1500mg/L NMP wastewater within 48h using this strain. Such a high mineralization degree indicated that the strain possessed a relatively complete NMP degradation pathway. In addition, the strain could completely degrade NMP with a concentration as high as 15000mg/L, which displayed the highest NMP degradation concentration with a high mineralization degree (63.2%) compared to the publicly available literatures. Finally, 10000mg/L NMP wastewater was treated by Burkholderia contaminans NCSL-HH10 and activated sludge under open environment, respectively. It was found that 95.7% NMP and 76.5% TOC were removed within 60h by NCSL-HH10, which was significantly higher than activated sludge (only 39.0% NMP and 30.2% TOC were removed within 84h).
李宁宁, 吕海慧, 于得水, 李雨桐, 王晔, 康建明, 薛宇昕, 金艳, 韩昫身, 于建国. N-甲基吡咯烷酮(NMP)高效降解菌筛选及应用研究[J]. 中国环境科学, 2025, 45(4): 2305-2313.
LI Ning-ning, Lü Hai-hui, YU De-shui, LI Yu-tong, WANG Ye, KANG Jian-ming, XUE Yu-xin, JIN Yan, HAN Xu-shen, YU Jian-guo. Screening and application of highly-efficient degrading strain for N-methyl-2-pyrrolidone (NMP). CHINA ENVIRONMENTAL SCIENCECE, 2025, 45(4): 2305-2313.
[1] Benhabbour S R, Kovarova M, Jones C, et al. Ultra-long-acting tunable biodegradable and removable controlled release implants for drug delivery[J]. Nature Communications, 2019,10(1):4324. [2] SönmezÖ, YıldızÖ,Çakır MÖ, et al. Influence of the addition of various ionic liquids on coal extraction with NMP[J]. Fuel, 2018,212:12-18. [3] Zeng W H, Li B B, Li H, et al. Mass produced NaA zeolite membranes for pervaporative recycling of spent N-Methyl-2-Pyrrolidone in the manufacturing process for lithium-ion battery[J]. Separation and Purification Technology, 2019,228:115741. [4] Zhou L, Zhang H C, Ahmad A L, et al. Hierarchical structure design of electrospun membrane for enhanced membrane distillation treatment of shrimp aquaculture wastewater[J]. Separation and Purification Technology, 2023,306:122591. [5] 于建国,孙庆,裘晟波,等.支撑国家新能源战略发展的锂资源开发[J].无机盐工业, 2023,55(1):1-14. Yu J G, Sun Q, Qiu S B, et al. Lithium resources development supporting national new energy strategy development[J]. Inorganic Chemicals Industry, 2023,55(1):1-14. [6] 何婷,孔娇,崔景植,等.退役锂离子电池电化学还原浸出及热力学研究[J].无机盐工业, 2022,54(12):34-43. He T, Kong J, Cui J, et al. Study on leaching and thermodynamic of spent lithium-ion batteries with electrochemical reduction[J]. Inorganic Chemicals Industry, 2022,54(12):34-43. [7] Ni H C, Arslan M, Liang Z S, et al. Mixotrophic denitrification processes in basalt fiber bio-carriers drive effective treatment of low carbon/nitrogen lithium slurry wastewater[J]. Bioresource Technology, 2022,364:128036. [8] 徐治伟,申魁文,马腾越,等.锂离子电池废弃正极浆料回收废水处理工程实例[J].水处理技术, 2023,49(11):147-149,156. Xu Z W, Shen K W, Ma T Y, et al. Research and design of wastewater treatment process for waste cathode slurry recycling of Li-ion batteries[J]. Technology of Water Treatment, 2023,49(11):147-149, 156. [9] Yoo K, Shin S M, Yang D H, et al. Biological treatment of wastewater produced during recycling of spent lithium primary battery[J]. Minerals Engineering, 2010,23(3):219-224. [10] Zhang X Y, Chen D, Jiang N, et al. New insights into algal-bacterial sludge granulation based on the tightly-bound extracellular polymeric substances regulation in response to N-Methylpyrrolidone[J]. Water Research, 2024,257:121754. [11] Bhojani G, Jani S, Saha N K. Facile biodegradation of N, N-dimethylformamide, N, N-dimethylacetamide and N-methyl-2-pyrrolidone by source-derived Bacillus strain APS1 for water reclamation and reuse[J]. Journal of Cleaner Production, 2022,334:130098. [12] Sherwood J, Farmer T J, Clark J H. Catalyst:Possible consequences of the N-Methyl Pyrrolidone REACH restriction[J]. Chem, 2018, 4(9):2010-2012. [13] 黄莉婷,韩昫身,金艳,等.煤化工反渗透浓水的高效降解菌株筛选、鉴定及应用研究[J].化工学报, 2021,72(9):4881-4891. Huang L T, Han X S, Jin Y, et al. Isolation, identification and application of highly efficient halotolerant strains for coal chemical reverse osmosis concentrate treatment[J]. CIESC Journal, 2021,72(9):4881-4891. [14] Chen S Y, Wang X Y, Shi X Y, et al. Integrated system of electro-catalytic oxidation and microbial fuel cells for the treatment of recalcitrant wastewater[J]. Chemosphere, 2024,354:141754. [15] Wu J J, Muruganandham M, Chang L T, et al. Ozone-based advanced oxidation processes for the decomposition of N-methyl-2-pyrolidone in aqueous medium[J]. Ozone:Science and Engineering, 2007,29(3):177-183. [16] Zolfaghari A, Mortaheb H R, Meshkini F. Removal of N-methyl-2-pyrrolidone by photocatalytic degradation in a batch reactor[J]. Industrial& Engineering Chemistry Research, 2011,50(16):9569-9576. [17] Kumar P, Verma S, Kaur R, et al. Enhanced photo-degradation of N-methyl-2-pyrrolidone (NMP):Influence of matrix components, kinetic study and artificial neural network modelling[J]. Journal of Hazardous Materials, 2022,434:128807. [18] Loh C H, Wu B, Ge L Y, et al. High-strength N-methyl-2-pyrrolidone-containing process wastewater treatment using sequencing batch reactor and membrane bioreactor:A feasibility study[J]. Chemosphere, 2018,194:534-542. [19] 于建国,韩昫身,金艳.页岩气压裂返排液生物处理技术研究进展[J].石油与天然气化工, 2022,51(5):131-138. Yu J G, Han X S, Jin Y. Biological treatment of shale gas flowback and produced water:a review[J]. Chemical Engineering of Oil&Gas, 2022, 51(5):131-138. [20] 田锐,陈威,王宗平,等.锂电池生产废水处理及中水回用工程实例[J].水处理技术, 2019,45(6):127-130. Tian R, Chen W, Wang Z P, et al. Engineering project of Lithium battery production wastewater treatment and reuse[J]. Technology of Water Treatment, 2019,45(6):127-130. [21] Herrero M, Stuckey D C. Bioaugmentation and its application in wastewater treatment:A review[J]. Chemosphere, 2015,140:119-128. [22] Wang J, Liu X L, Jiang X B, et al. Nitrate stimulation of N-Methylpyrrolidone biodegradation by Paracoccus pantotrophus:Metabolite mechanism and Genomic characterization[J]. Bioresource Technology, 2019,294:122185. [23] Wang J, Chi Q, Zhang R R, et al. Evaluation of N-methylpyrrolidone bio-mineralization mechanism and bacterial community evolution under denitrification environment[J]. Journal of Cleaner Production, 2022,343:130945. [24] Cai S, Cai T M, Liu S Y, et al. Biodegradation of N-Methylpyrrolidone by Paracoccus sp. NMD-4and its degradation pathway[J]. International Biodeterioration& Biodegradation, 2014, 93:70-77. [25] Gao Y Y, Chen T, Hou Y F, et al. The roles of Methylobacterium organophilum and Sphingomonas melonis for accelerating N-methyl pyrrolidone (NMP) biodegradation[J]. Journal of Water Process Engineering, 2023,56:104327. [26] Li N N, Han X S, Lv H H, et al. Biodegradation of N-methyl-2-pyrrolidone (NMP) in wastewater:A review of current knowledge and future perspectives[J]. Journal of Cleaner Production, 2025,486:144452. [27] Jeon B Y, Yi J Y, Park D H. Estimation on metabolic pathway of Pseudomonas sp. SMIC-3for 1-methyl-2-pyrrolidinone based on physiological and biochemical analyses[J]. Korean Journal of Chemical Engineering, 2014,31:475-484. [28] Tang R, Han X S, Jin Y, et al. Do increased organic loading rates accelerate aerobic granulation in hypersaline environment?[J]. Journal of Environmental Chemical Engineering, 2022,10(6):108775. [29] Kumar S, Stecher G, Li M, et al. MEGA X:Molecular Evolutionary Genetics Analysis across Computing Platforms[J]. Molecular Biology and Evolution, 2018,35(6):1547-1549. [30] Tamura K, Peterson D, Peterson N, et al. MEGA5:molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods[J]. Molecular Biology and Evolution, 2011,28(10):2731-2739. [31] Thompson J D, Higgins D G, Gibson T J. CLUSTAL W:Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice[J]. Nucleic Acids Research, 1994,22(22):4673-4680. [32] O'Sullivan L A, Mahenthiralingam E. Biotechnological potential within the genus Burkholderia[J]. Letters in Applied Microbiology, 2005,41(1):8-11. [33] Kumari S, Das S. Bacterial enzymatic degradation of recalcitrant organic pollutants:Catabolic pathways and genetic regulations[J]. Environmental Science and Pollution Research, 2023,30(33):79676-79705. [34] Chen S Y, Zhu X H, Zhu G, et al. N-methyl pyrrolidone manufacturing wastewater as the electron donor for denitrification:From bench to pilot scale[J]. Science of the Total Environment, 2024,912:169517. [35] 楼超楠,韩昫身,金艳,等.不同碳源对微生物反硝化性能的影响[J].华东理工大学学报(自然科学版), 2023,49(5):660-669. Lou C N, Han X S, Jin Y, et al. Effect of different carbon sources on microbial denitrification performance[J]. Journal of East China University of Science and Technology, 2023,49(5):660-669. [36] Daniel R M, Peterson M E, Danson M J, et al. The molecular basis of the effect of temperature on enzyme activity[J]. Biochemical Journal, 2010,425(2):353-360. [37] Campbell H L, Striebig B A. Evaluation of N-methylpyrrolidone and its oxidative products toxicity utilizing the microtox assay[J]. Environmental Science& Technology, 1999,33(11):1926-1930. [38] 张文涛,胡友彪,高良敏,等.甲胺废水资源化探究[J].环境科学与技术, 2011,34(1):180-183. Zhang W T, Hu Y B, Gao L M, et al. Reutilization ofmethylamine wastewater[J]. Environmental Science& Technology, 2011,34(1):180-183. [39] Han X S, Jin Y, Yu J G. Rapid formation of aerobic granular sludge by bioaugmentation technology:A review[J]. Chemical Engineering Journal, 2022,437:134971. [40] Han X S, Huang L T, Jin Y, et al. Bioaugmentation treatment of coal chemical reverse osmosis concentrate in biological contact oxidation system coupled with ozone pretreatment[J]. Journal of Cleaner Production, 2023,428:139455.
