Abstract：In this research, a strain of bacteria with vanadium reduction ability was isolated from the vanadium contaminated soil, and the influence of different V(V) concentration, inoculum amount and pH value on V(V) reduction was investigated. Meanwhile, the extracellular and intracellular V(V) reduction by strain and the enzyme activity changes involved in V(V) reduction were characterized to reveal the mechanism of V(V) bioreduction. The results showed that strain NC1-2 was identified as a strain of Enterobacter kobei. When initial V(V) concentration was 160mg/L, the reduction rate of V(V) by the strain was 96.29%. Increase bacterial inoculation accelerated V(V) reduction by the strain, and pH 8.0 was the optimal condition. With decreased the cell membrane permeability, the reduction rate of V(V) increased from 71.2% to 75.0%. There were differences in the reduction of V(V) by different subcellular fractions. It was found that the reduction rates of V(V) by extracellular exudates and cytoplasmic fractions were 41.71% and 80.17%, respectively; No V(V) reduction occurred in the cell membrane fraction. The nitrite reductase (NIR) activity and reduced nicotinamide adenine dinucleotide (NADH) content were increased to different degrees during the reduction of V(V) by the strains. Fourier transform infrared spectroscopy (FTIR) analysis showed the hydroxyl, carbonyl and amide groups on the surface of bacteria were involved in biosorption. Scanning electron microscope (SEM) showed the precipitation around the bacterial body with V(V) reduction. Energy dispersive spectroscopy (EDS) indicated the presence of vanadium in the precipitate. X-ray photoelectron spectroscopy (XPS) analysis showed that strain NC1-2 reduced V(V) to V(IV). Transmission electron microscope (TEM) results indicated that vanadium was precipitated within the bacteria. In summary, the results suggested that strain NC1-2 was able to reduce V(V) and form insoluble V(IV) precipitates intracellularly and extracellularly, and NIR and NADH were involved in the intracellular biotransformation. The research revealed the characteristics and intrinsic mechanism of V(V) reduction by Enterobacter kobei NC1-2, and preliminarily analyzed its intracellular electron transfer. The isolated strain has good application prospects in vanadium pollution remediation.
周睿, 周雅琪, 王丽丽, 盛米雪, 司友斌. 一株钒还原菌的分离鉴定及V(V)还原机理研究[J]. 中国环境科学, 2023, 43(6): 2926-2937.
ZHOU Rui, ZHOU Ya-qi, WANG Li-li, SHENG Mi-xue, SI You-bin. Isolation and identification of a vanadium reducing bacterium and the mechanism of V(V) bioreduction. CHINA ENVIRONMENTAL SCIENCECE, 2023, 43(6): 2926-2937.
Huang J H, Huang F, Evans L, et al. Vanadium:Global (bio) geochemistry[J]. Chemical Geology, 2015,417:68-89.
[2]
Long Z J, Huang Y, Zhang W, et al. Effect of different industrial activities on soil heavy metal pollution, ecological risk, and health risk[J]. Environmental Monitoring and Assessment, 2021,193(1):20.
[3]
Vasseghian Y, Sadeghi S, Vilas-Boas J A, et al. A global systematic review, meta-analysis, and risk assessment of the concentration of vanadium in drinking water resources[J]. Chemosphere, 2021,267:128904.
[4]
Yang J, Teng Y G, Wu J, et al. Current status and associated human health risk of vanadium in soil in China[J]. Chemosphere, 2017,171:635-643.
[5]
李 波,喻庆国,危 锋,等.滇西北剑湖沉积物钒空间分布特征和生态风险[J]. 中国环境科学, 2019,39(5):2219-2229. Li B, Yu G Q, Wei F, et al. Spatial distribution characteristics and ecological risks of vanadium in sediments from Jianhu Lake[J]. China Environmental Science, 2019,39(5):2219-2229.
[6]
Zhang B G, Wang S, Diao M H, et al. Microbial community responses to vanadium distributions in mining geological environments and bioremediation assessment[J]. Journal of Geophysical Research:Biogeosciences, 2019,124(3):601-615.
[7]
郭 威,殷淑华,徐建新,等.三峡库区(重庆-宜昌段)沉积物中钒的污染特征及生态风险评价[J]. 环境科学, 2016,37(9):3333-3339. Guo W, Yin S H, Xu X J, et al. Pollution characteristics and ecological risk assessment of vanadium in sediments of the Three Gorges Reservoir (Chongqing-Yichang Section)[J]. Environment Science, 2016,37(9):3333-3339.
[8]
Ortiz-Bernad I, Anderson R T, Vrionis H A, et al. Vanadium respiration by Geobacter metallireducens:Novel strategy for in situ removal of vanadium from groundwater[J]. Applied and Environmental Microbiology, 2004,70(5):3091.
[9]
李亚瑾,孙志高,李 晓,等.闽江河口芦苇与短叶茳芏空间扩展对湿地植物钒生物富集的影响[J]. 环境科学学报, 2020,40(12):4559-4569. Li Y J, Sun Z G, Li X, et al. Effects of spatial expansion between Phragmites australis and Cyperus malaccensis on bioaccumulation of vanadium in plants of different marshes in the min river estuary[J]. Acta Scientiae Circumstantiae, 2020,40(12):4559-4569.
[10]
Chen L, Liu J R, Hu W F, et al. Vanadium in soil-plant system:Source, fate, toxicity, and bioremediation[J]. Journal of Hazardous Materials, 2021,405:124200.
[11]
Aihemaiti A, Gao Y, Meng Y, et al. Review of plant-vanadium physiological interactions, bioaccumulation, and bioremediation of vanadium-contaminated sites[J]. Science of the Total Environment, 2020,712:135637.
[12]
Gustafsson J P. Vanadium geochemistry in the biogeosphere -speciation, solid-solution interactions, and ecotoxicity[J]. Applied Geochemistry, 2019,102:1-25.
[13]
Imtiaz M, Rizwan M S, Xiong S, et al. Vanadium, recent advancements and research prospects:A review[J]. Environment International, 2015,80:79-88.
[14]
Wanty R B, Goldhaber M B. Thermodynamics and kinetics of reactions involving vanadium in natural systems:Accumulation of vanadium in sedimentary rocks[J]. Geochimica et Cosmochimica Acta, 1992,56(4):1471-1483.
[15]
郑家传,张建荣,刘希雯,等.污染场地六价铬的还原和微生物稳定化研究[J]. 环境科学, 2014,35(10):3882-3887. Zheng J C, Zhang J R, Liu X W, et al. Bioremediation of chromium (Ⅵ) contaminated site by reduction and microbial stabilization of chromium[J]. Environment Science, 2014,35(10):3882-3887.
[16]
谷 倩,刘 欢,张宝刚,等.钒污染土壤地下水的修复技术研究进展[J]. 地球科学, 2018,43(S1):84-96. Gu Q, Liu H, Zhang B G, et al. Research progress on remediation technology of vanadium contaminated soil and groundwater[J]. Earth Science, 2018,43(S1):84-96.
[17]
王鑫伟,孙明明,朱国繁,等.钒污染土壤生物修复研究进展[J]. 土壤, 2020,52(5):873-882. Wang W X, Sun M M, Zhu G F, et al. Research progresses of vanadium-contaminated soil bioremediation[J]. Soils, 2020,52(5):873-882.
[18]
Wang G, Zhang B, Li S, et al. Simultaneous microbial reduction of vanadium (V) and chromium (VI) by Shewanella loihica PV-4[J]. Bioresource Technology, 2017,227:353-358.
[19]
Yelton A P, Williams K H, Fournelle J, et al. Vanadate and acetate biostimulation of contaminated sediments decreases diversity, selects for specific taxa, and decreases aqueous V5+ concentration[J]. Environmental Science & Technology, 2013,47(12):6500-6509.
[20]
Antipov A N, Lyalikova N N, Khijniak T V, et al. Vanadate reduction by molybdenum-free dissimilatory nitrate reductases from vanadate-reducing bacteria[J]. IUBMB Life, 2000,50(1):39-42.
[21]
Antipov, A N, Lyalikova N N, L'Vov N P. Vanadium-binding protein excreted by vanadate-reducing bacteria[J]. IUBMB Life, 2000,49(2):137-141.
[22]
Ueki T, Yamaguchi N, Romaidi, et al. Vanadium accumulation in ascidians:A system overview[J]. Coordination Chemistry Reviews, 2015,301-302:300-308.
[23]
Li J L, Zhang B G. Woodchip-sulfur packed biological permeable reactive barrier for mixotrophic vanadium (V) detoxification in groundwater[J]. Science China Technological Sciences, 2020,63(11):2283-2291.
[24]
Sun X, Qiu L, Kolton M, et al. V(V) reduction by Polaromonas spp. in vanadium mine tailings[J]. Environmental Science & Technology, 2020,54(22):14442-14454.
[25]
Carpentier W, De S L, Van B J, et al. Respiration and growth of Shewanella oneidensis MR-1using vanadate as the sole electron acceptor[J]. Journal of Bacteriology, 2005,187(10):3293.
[26]
李 劲,杨玉蓉,朱广森,等.Shewanella oneidensis MR-1和Shewanella putrefaciens对V(Ⅴ)的还原及其影响因素[J]. 中国环境科学, 2020,40(1):414-421. Li J, Yang Y R, Zhu G S, et al. The reduction of V(V) by Shewanella oneidensis MR-1and Shewanella putrefaciens and influencing factors[J]. China Environmental Science, 2020,40(1):414-421.
[27]
Zhang B, Cheng Y, Shi J, et al. Insights into interactions between vanadium (V) bio-reduction and pentachlorophenol dechlorination in synthetic groundwater[J]. Chemical Engineering Journal, 2019,375:121965.
[28]
郝丽婷.钒污染地下水的微生物修复与机理研究[D]. 北京:中国地质大学, 2019:24-27. Hao L T. Microbial remediation and mechanism study of vanadium contaminated groundwater[D]. Beijing:China University of Geosciences (Beijing), 2019:24-27.
[29]
Farmer J J, Fanning G R, Davis B R, et al. Escherichia fergusonii and Enterobacter taylorae, two new species of Enterobacteriaceae isolated from clinical specimens[J]. Journal of Clinical Microbiology, 1985,21(1):77-81.
[30]
Tamura K, Sakazaki R, Kosako Y, et al. Leclercia adecarboxylata Gen. Nov., Comb. Nov., formerly known as Escherichia adecarboxylata [J]. Current Microbiology, 1986,13(4):179-184.
[31]
Kosako Y, Tamura K, Sakazaki R, et al. Enterobacter kobei sp. nov., a new species of the family Enterobacteriaceae resembling Enterobacter cloacae [J]. Current Microbiology, 1996,33(4):261-265.
[32]
Zhu Y, Yan J, Xia L, et al. Mechanisms of Cr(VI) reduction by Bacillus sp. CRB-1, a novel Cr(VI)-reducing bacterium isolated from tannery activated sludge[J]. Ecotoxicology and Environmental Safety, 2019,186:109792.
[33]
Zhang J, Dong H, Zhao L, et al. Microbial reduction and precipitation of vanadium by mesophilic and thermophilic methanogens[J]. Chemical Geology, 2014,370:29-39.
[34]
Zhang B, Li Y, Fei Y, et al. Novel pathway for Vanadium(V) bio-detoxification by gram-positive Lactococcus raffinolactis [J]. Environmental Science & Technology, 2021,55(3):2121-2131.
[35]
Xue Y, Guo J, Lian J, et al. Effects of a higher hydraulic shear force on denitrification granulation in upflow anoxic sludge blanket reactors[J]. Biochemical Engineering Journal, 2016,105:136-143.
[36]
Filik H, Berker K I, Balkis N, et al. Simultaneous preconcentration of vanadium(V/IV) species with palmitoyl quinolin-8-ol bonded to amberlite XAD 2and their separate spectrophotometric determination with 4-(2-pyridylazo)-resorcinol using CDTA as masking agent[J]. Analytica Chimica Acta, 2004,518(1):173-179.
[37]
van Marwijk J, Opperman D J, Piater L A, et al. Reduction of vanadium(V) by Enterobacter cloacae EV-SA01isolated from a South African deep gold mine[J]. Biotechnology Letters, 2009,31(6):845-849.
[38]
Lai C Y, Dong Q Y, Chen J X, et al. Role of extracellular polymeric substances in a methane based membrane biofilm reactor reducing vanadate[J]. Environmental Science & Technology, 2018,52(18):10680-10688.
[39]
Shi J, Zhang B, Cheng Y, et al. Microbial vanadate reduction coupled to co-metabolic phenanthrene biodegradation in groundwater[J]. Water Research, 2020,186:116354.
[40]
Xu X, Xia S, Zhou L, et al. Bioreduction of vanadium (V) in groundwater by autohydrogentrophic bacteria:Mechanisms and microorganisms[J]. Journal of Environmental Sciences, 2015,30:122-128.
[41]
Antipov A N, Khijniak T V. Vanadate reduction under alkaline conditions by haloalkaliphilic Halomonas strains[J]. Microbiology, 2016,85(6):658-663.
[42]
Costa P J, Garribba E, Santos M F A, et al. Vanadium and proteins:Uptake, transport, structure, activity and function[J]. Coordination Chemistry Reviews, 2015,301-302:49-86.
[43]
Ghazvini P T M, Mashkani S G. Effect of salinity on vanadate biosorption by Halomonas sp. GT-83:Preliminary investigation on biosorption by micro-PIXE technique[J]. Bioresource Technology, 2009,100(8):2361-2368.
[44]
Zhang B, Hao L, Tian C, et al. Microbial reduction and precipitation of vanadium (V) in groundwater by immobilized mixed anaerobic culture[J]. Bioresource Technology, 2015,192:410-417.
[45]
Li L, Zhang B, Li L, et al. Microbial selenate detoxification linked to elemental sulfur oxidation:Independent and synergic pathways[J]. Journal of Hazardous Materials, 2022,422:126932.
[46]
Zhang B, Wang Z, Shi J, et al. Sulfur-based mixotrophic bio-reduction for efficient removal of chromium (VI) in groundwater[J]. Geochimica et Cosmochimica Acta, 2020,268:296-309.
[47]
Shi J, Li Z, Zhang B, et al. Synergy between pyridine anaerobic mineralization and vanadium (V) oxyanion bio-reduction for aquifer remediation[J]. Journal of Hazardous Materials, 2021,418:126339.
[48]
Mendz G L. Redox and specific effects of vanadium ions on respiratory enzymes[J]. Biometals, 1999,12(1):35-45.
