寡养单胞菌对S~(2-)氧化特性及主要代谢途径
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摘要
以无机硫(S2)-为目标污染物,从北京东沙河黑臭水体筛选出一株能够高效氧化S~(2-)的土著硫氧化菌,并对其进行16S rRNA测序及生理生化特性鉴定,结果表明,该菌株属于寡养单胞菌属(Stenotrophomonas),命名为Stenotrophomonas sp.sp3.菌株sp3在温度25℃,初始pH值7.0,初始葡萄糖浓度0.25%,初始菌浓度1.00g/L时,菌株sp3对S~(2-)的氧化率最高.在此适宜条件下,反应60h后,菌株sp3对S~(2-)的最高氧化率达到86.6%.S~(2-)的浓度在整个氧化过程中持续下降后保持稳定,在此过程中产生了S~0,S_2O_3~(2-),SO_3~(2-)以及SO_4~(2-)这4种其它存在形态的硫.随着S~(2-)被氧化,SO_4~(2-)的浓度呈缓慢上升的趋势.利用高通量测序技术推测菌株sp3主要是通过副球菌硫氧化途径(PSO)将非稳定态的S~(2-)逐步转化为稳定态的SO_4~(2-).在此代谢途径中,一部分S~(2-)被氧化为S~0,另一部分S~(2-)则直接氧化为SO_3~(2-);S~0与SO_3~(2-)可自发反应生成S_2O_3~(2-),而S_2O_3~(2-)发生歧化反应再释放出SO_3~(2-)和S~0;反应生成的部分SO_3~(2-)继续直接氧化为SO_4~(2-).菌株sp3在黑臭水体的水质净化过程中有一定的应用前景.
Inorganic sulfur(S~(2-)) was used as the target pollutant, and a single strain capable of S~(2-)-oxidizing was obtained from Dongsha river(a black-stinking river) located in Beijing. Through both of the physiological-biochemical characteristics and the 16 S rRNA sequencing experiment, this single strain was identified as Stenotrophomonas, and named as Stenotrophomonas sp.sp3. The experiment on the growth and S~(2-)-oxidizing characteristics of this strain sp3 was then carried out. The best conditions of S~(2-)-oxidizing were as follows: initial pH of 7.0, 25℃, initial glucose concentration of 0.25% along with initial cell concentration of 1.00 g/L. Under the above bio-oxidation conditions, the highest S~(2-)-oxidizing ratio was arrived at 86.6% by this strain sp3 after the bio-reaction of 60 h. The concentration of S~(2-)decreased continually and then kept in a stable status in the whole bio-reaction process, and other sulphur chemical forms including S~0, S_2O_3~(2-), SO_3~(2-)and SO_4~(2-)were produced in this period. With the bio-oxidation of S~(2-), the concentration of SO_4~(2-)increased slowly. High-throughput sequencing technologies were used to investigate the main metabolic pathway of S~(2-)bio-oxidation by this strain sp3. The paracoccus sulfur oxidation process was the possibly main metabolic pathway of S~(2-)(unstable valence state) to SO_4~(2-)(stable valence state). It was initially concluded that a portion of S~(2-)was oxidized to S~0, while other part of S~(2-)was directly oxidized to SO_3~2; and then S~0 reacted with SO_3~(2-)spontaneously to form S_2O_3~(2-), while the latter was released SO_3~(2-)and S~0 by the disproportionate reaction again. The SO_3~(2-)accumulated gradually in the system was further directly oxidized to SO_4~(2-). This study indicated that this strain sp3 has potential application in water purification of the black-stinking wate bodies.
Inorganic sulfur(S~(2-)) was used as the target pollutant, and a single strain capable of S~(2-)-oxidizing was obtained from Dongsha river(a black-stinking river) located in Beijing. Through both of the physiological-biochemical characteristics and the 16 S rRNA sequencing experiment, this single strain was identified as Stenotrophomonas, and named as Stenotrophomonas sp.sp3. The experiment on the growth and S~(2-)-oxidizing characteristics of this strain sp3 was then carried out. The best conditions of S~(2-)-oxidizing were as follows: initial pH of 7.0, 25℃, initial glucose concentration of 0.25% along with initial cell concentration of 1.00 g/L. Under the above bio-oxidation conditions, the highest S~(2-)-oxidizing ratio was arrived at 86.6% by this strain sp3 after the bio-reaction of 60 h. The concentration of S~(2-)decreased continually and then kept in a stable status in the whole bio-reaction process, and other sulphur chemical forms including S~0, S_2O_3~(2-), SO_3~(2-)and SO_4~(2-)were produced in this period. With the bio-oxidation of S~(2-), the concentration of SO_4~(2-)increased slowly. High-throughput sequencing technologies were used to investigate the main metabolic pathway of S~(2-)bio-oxidation by this strain sp3. The paracoccus sulfur oxidation process was the possibly main metabolic pathway of S~(2-)(unstable valence state) to SO_4~(2-)(stable valence state). It was initially concluded that a portion of S~(2-)was oxidized to S~0, while other part of S~(2-)was directly oxidized to SO_3~2; and then S~0 reacted with SO_3~(2-)spontaneously to form S_2O_3~(2-), while the latter was released SO_3~(2-)and S~0 by the disproportionate reaction again. The SO_3~(2-)accumulated gradually in the system was further directly oxidized to SO_4~(2-). This study indicated that this strain sp3 has potential application in water purification of the black-stinking wate bodies.
引文
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[2]住房和城乡建设部和环境保护部.全国城市黑臭水体整治信息发布平台[EB/OL]. http://www.hcstzz.com/2017-08.Ministry of Housing and Urban-Rural Construction and Ministry of Environmental Protection. National information publishing platform for urban black and odorous water treatment[EB/OL]. http://www.hcstzz.com/2017-08.
[3] Song C, Liu X L, Song Y H, et al. Key blackening and stinking pollutants in Dongsha River of Beijing:Spatial distribution and source identification[J]. Journal of Environmental Management, 2017,200:335-346.
[4] Tang W, Shan B, Zhang H, et al. Heavy metal contamination in the surface sediments of representative limnetic ecosystems in eastern China[J]. Scientific Reports, 2014,4:7152-7158.
[5]王玉琳,汪靓,华祖林.黑臭水体中不同浓度Fe2+、S2-与DO和水动力关系[J].中国环境科学, 2018,38(2):627-633.Wang Y L, Wang L, Hua Z L. The relationships of different concentration Fe2+, S2-with hydrodynamics, DO in black bloom water based on quantile regression method[J]. China Environmental Science,2018,38(2):627-633.
[6]王旭,王永刚,孙长虹,等.城市黑臭水体形成机理与评价方法研究进展[J].应用生态学报, 2016,27(4):1331-1340.Wang X, Wang Y G, Sun C H, et al. Formation mechanism and assessment method for urban black and odorous water body:A review[J]. Chinese Journal of Applied Ecology, 2016,27(4):1331-1340.
[7] Liu C, Shen Q, Zhou Q, et al. Precontrol of algae-induced black blooms through sediment dredging at appropriate depth in a typical eutrophic shallow lake[J]. Ecological Engineering, 2015,77:139-145.
[8] Jiang Y, Xie P, Nie Y. Concentration and bioaccumulation of cyanobacterial bioactive and odorous metabolites occurred in a large,shallow Chinese lake[J]. Bulletin of Environmental Contamination and Toxicology, 2014,93:643-648.
[9] Sheng Y, Qu Y, Ding C, et al. A combined application of different engineering and biological techniques to remediate a heavily polluted river[J]. Ecological Engineering, 2013,57(57):1-7.
[10] Gu D G, Xu H, He Y, et al. Remediation of urban river water by Pontederia cordata combined with artificial aeration:Organic matter and nutrients removal and root-adhered bacterial communities[J].International Journal of Phytoremediation, 2015,17(11):1105-1114.
[11] He W, Shang J G, Lu X, et al. Effects of sludge dredging on the prevention and control of algae-caused black bloom in Taihu Lake,China[J]. Journal of Environmental Sciences, 2013,25(3):430-440.
[12]陈超,钟继承,范成新,等.疏浚对湖泛的影响:以太湖八房港和闾江口水域为例[J].中国环境科学, 2014,34(8):2071-2077.Chen C, Zhong J C, Fan C X, et al. Effects of sludge dredging on black bloom:a case study of Bafang port and Lujiang port of Taihu Lake[J]. China Environmental Science, 2014,34(8):2071-2077.
[13]刘树娟,陈磊,钟润生,等.硝酸钙对河流底泥中含硫化合物嗅味原位控制[J].环境科学研究, 2012,25(6):691-698.Liu S J, Chen L, Zhong R S, et al. In situ control of odor in sulfide-containing compounds with calcium nitrate in river sediments[J]. Research of Environmental Sciences, 2012,25(6):691-698.
[14] Liang-Liang T, Peng L I, Jing-Min D, et al. The Chemical Coagulation-Advanced Oxidation Composite Process for Treating Sulfur-contained Tannery Wastewater[J]. Leather&Chemicals, 2011,28(6):8-10.
[15] Wang M, Yang G, Min H, et al. Bioaugmentation with the nicotine-degrading bacterium Pseudomonas sp. HF-1in a sequencing batch reactor treating tobacco wastewater:Degradation study and analysis of its mechanisms[J]. Water Research, 2009,43(17):4187-4196.
[16] Jin M, Wang X W, Gong T S, et al. A novel membrane bioreactor enhanced by effective microorganisms for the treatment of domestic wastewater[J]. Applied Microbiology&Biotechnology, 2005,69(2):229-235.
[17] Chen J, Koopman B, Fang G, et al. Bioaugmentation with Gordonia strain JW8 in treatment of pulp and paper wastewater[J]. Clean Technologies&Environmental Policy, 2012,14(5):899-904.
[18] Semrany S, Favier L, Djelal H, et al. Bioaugmentation:Possible solution in the treatment of Bio-Refractory Organic Compounds(Bio-ROCs)[J]. Biochemical Engineering Journal, 2012,69(51):75-86.
[19]徐熊鲲,谢翼飞,陈政阳,等.曝气强化微生物功能菌修复黑臭水体[J].环境工程学报, 2017,11(8):4559-4565.Xu X K, Xie Y F, Chen Z Y, et al. Remediation of urban black-odorous river by aeration enhancing functional strains[J]. Chinese Journal of Environmental Engineering, 2017,11(8):4559-4565.
[20]吴霞,谢悦波.直接投菌法在城市重污染河流治理中的应用研究[J].环境工程学报, 2014,8(8):3331-3336.Wu X, Xie Y B. Application of directly adding microbial agent to improve water quality of heavily polluted urban river[J]. Chinese Journal of Environmental Engineering, 2014,8(8):3331-3336.
[21] Guo H, Chen C, Lee D J, et al. Proteomic analysis of sulfur–nitrogen-carbon removal by Pseudomonas sp. C27under micro–aeration condition[J]. Enzyme and Microbial Technology, 2014,56:20-27.
[22]东秀珠,蔡妙英.常见细菌系统鉴定乎册[M].北京:科学出版社,2002.Dong X Z, Cai M Y. Systematic identification of common bacteria[M].Beijing:Science Press, 2002.
[23]沈萍,范秀容,李广武.微生物学实验[M].北京:高等教育出版社,2003.Shen P, Fan X R, Li G W. Microbiology Experiment[M]. Beijing:Higher Education Press, 2003.
[24] Zhuang R, Lou Y, Qiu X, et al. Identification of a yeast strain able to oxidize and remove sulfide high efficiently[J]. Applied Microbiology&Biotechnology, 2017,101(1):391-400.
[25]国家环境保护总局《水和废水监测分析方法》编委会.水和废水监测分析方法[M]. 4版.中国环境科学出版社, 2002.Editorial board of water and wastewater monitoring and analysis method of the State Environmental Protection Administration. Water and Wastewater Monitoring and Analysis Method(4)[M]. China Environmental Science Press, 2002.
[26]冯玉雪,毛缜,吕蒙蒙.一株DDT降解菌的筛选及其降解特性[J].中国环境科学, 2018,(5):1935-1942.Feng Y X, Mao Z, LüM M. Screening and degradation characteristics of a DDT-degrading bacteria[J]. China Environmental Science,2018(5):1935-1942.
[27] Lambert T, MarieCécile P, Fran?ois D, et al. Characterization of the Chromosomal aac(6′)-Iz Gene of Stenotrophomonas maltophilia[J].Antimicrob Agents Chemother, 1999,43(10):2366-2371.
[28] Shen Y J, Lu P, Mei H, et al. Isolation of a methyl parathion-degrading strain Stenotrophomonas sp. SMSP-1and cloning of the ophc2gene[J].Biodegradation, 2010,21(5):785-792.
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