Aspergillus tubingensis介导植酸盐水解促进U(VI)-PO_4~(3-)生物矿化
|
摘要
从广东某铀尾矿库水下沉积物中分离筛选出了一株能水解植酸盐的真菌M5-1,对其菌落形态、ITS序列、最适生长pH值、对铀的耐受性及其水解植酸盐的效果进行了分析,随后对M5-1生物矿化铀过程中pH值、正磷酸盐浓度、铀浓度、铀去除率的变化进行了监测,对矿化产物的主要元素和矿物组成进行了分析.证实了真菌M5-1为Aspergillustubingensis(MH978623),其最适生长pH值范围为6～7,对铀(～0.84mmol/L)具有较强的耐受性;Aspergillus tubingensis介导植酸盐水解促进U(Ⅵ)-PO_4~(3-)矿化62d后,铀的去除率达95.2%;Aspergillus tubingensis介导U(Ⅵ)-PO_4~(3-)矿化过程中可能形成了难溶的氢铀云母和变钠铀云母矿物.结果表明,Aspergillustubingensis能有效水解植酸盐释放可溶性正磷酸盐,从而促进U(Ⅵ)-PO_4~(3-)矿化.研究结果为采用Aspergillus tubingensis介导植酸盐水解原位修复铀污染地表水提供了试验依据.
The fungus M5-1 capable of hydrolyzing phytate was isolated from the sediments of a uranium tailings repository in Guangdong Province. Its colony morphology, ITS sequences, suitable growth pH value, tolerance against uranium and effect on hydrolysis of phytate were systematically studied. The variations of pH value, orthophosphate concentration, uranium concentration and removal efficiency of uranium were monitored. The main elements and mineral components of the biomineralization products were also analyzed. It was found that the strain M5-1 was Aspergillus tubingensis(MH978623) with optimal growth at pH 6～7, and with high tolerance against uranium(～0.84 mmol/L). After biomineralization of U(VI)-PO_4~(3-)promoted by Aspergillus tubingensis mediated phytate hydrolysis for 62 days, the removal efficiency of uranium reached 95.2%. SEM-EDS and XRD analyses indicated that the insoluble chernikovite and metanatroautunite were formed during the biomineralization of U(VI)-PO_4~(3-). The results showed that Aspergillus tubingensis could effectively hydrolyze phytate to release soluble orthophosphate, which promoted the mineralization of U(VI)-PO_4~(3-). These results provided an experimental basis for the in-situ bioremediation of uranium contaminated surface water by biomineralization of U(VI)-PO_4~(3-)promoted by Aspergillus tubingensis mediated phytate hydrolysis.
The fungus M5-1 capable of hydrolyzing phytate was isolated from the sediments of a uranium tailings repository in Guangdong Province. Its colony morphology, ITS sequences, suitable growth pH value, tolerance against uranium and effect on hydrolysis of phytate were systematically studied. The variations of pH value, orthophosphate concentration, uranium concentration and removal efficiency of uranium were monitored. The main elements and mineral components of the biomineralization products were also analyzed. It was found that the strain M5-1 was Aspergillus tubingensis(MH978623) with optimal growth at pH 6～7, and with high tolerance against uranium(～0.84 mmol/L). After biomineralization of U(VI)-PO_4~(3-)promoted by Aspergillus tubingensis mediated phytate hydrolysis for 62 days, the removal efficiency of uranium reached 95.2%. SEM-EDS and XRD analyses indicated that the insoluble chernikovite and metanatroautunite were formed during the biomineralization of U(VI)-PO_4~(3-). The results showed that Aspergillus tubingensis could effectively hydrolyze phytate to release soluble orthophosphate, which promoted the mineralization of U(VI)-PO_4~(3-). These results provided an experimental basis for the in-situ bioremediation of uranium contaminated surface water by biomineralization of U(VI)-PO_4~(3-)promoted by Aspergillus tubingensis mediated phytate hydrolysis.
引文
[1] Zheng X Y, Wang X Y, Shen Y H, et al. Biosorption and biomineralization of uranium(VI)by Saccharomyces cerevisiaecrystal formation of chernikovite[J]. Chemosphere, 2017,175:161-169
[2] Yuan F, Wu C, Cai Y, et al. Synthesis of phytic acid-decorated titanate nanotubes for high efficient and high selective removal ofU(VI)[J]. Chemical Engineering Journal, 2017,322:353-365
[3] Parrish R R, Horstwood M, Arnason J G, et al. Depleted uranium contamination by inhalation exposure and its detection after~20 years:implications for human health assessment[J]. Science of the Total Environment, 2008,390(1):58-68.
[4] Taylor D M, Taylor S K. Environmental uranium and human health[J].Reviews on Environmental Health, 1997,12(3):147-158.
[5] Newsome L,Morris K, Lloyd J R. The biogeochemistry and bioremediation of uranium and other priority radionuclides[J].Chemical Geology, 2014,363:164-184.
[6] Beazley M J, Martinez R J, Sobecky P A, et al. Uranium biomineralization as a result of bacterial phosphatase activity:insights from bacterial isolates from a contaminated subsurface[J].Environmental Science&Technology, 2007,41(16):5701-5707.
[7] Wu W, Carley J, Luo J, et al. In situ bioreduction of uranium(VI)to submicromolar levels and reoxidation by dissolved oxygen[J].Environmental Science&Technology, 2007,41(16):5716-5723.
[8] Moon H S, Komlos J, Jaffe P R. Biogenic U(IV)oxidation by dissolved oxygen and nitrate in sediment after prolonged U(VI)/Fe(Ⅲ)/SO42-reduction[J]. Journal of Contaminant Hydrology, 2009,105(1):18-27.
[9] Wan J, Tokunaga T K, Brodie E, et al. Reoxidation of bioreduced uranium under reducing conditions[J]. Environmental Science&Technology,2005,39(16):6162-6169.
[10] Wu W, Carley J, Fienen M, et al. Pilot-scale in situ bioremediation of uranium in a highly contaminated aquifer. 1. conditioning of a treatment zone[J]. Environmental Science&Technology, 2006,40(12):3978-3985.
[11] Wufuer R, Wei Y, Lin Q,et al. Uranium bioreduction and biomineralization[J]. Advances in Applied Microbiology, 2017,101:137-168.
[12] Knox A S,Brigmon R L, Kaplan D I, et al. Interactions among phosphate amendments, microbes and uranium mobility in contaminated sediments[J]. Science of the Total Environment,2008,395(2):63-71.
[13] Liang X,Hillier S, Pendlowski H, et al. Uranium phosphate biomineralization by fungi[J]. Environmental Microbiology, 2015,17(6):2064-2075.
[14] Salome K. Non-reductive biomineralization of U(Ⅵ)-phosphate minerals through the activities of microbial phytases[D]. Atlanta:Georgia Institute of Technology, 2013:48-52.
[15]王晓东,胡海滨,麦康森,等.土壤中单宁和植酸降解菌的筛选、鉴定及液态发酵研究[J].水产学报,2016,40(10):1634-1642.Wang X D, Hu H B, Mai K S, et al. Screening and identification of tannic acid and phytic acid degradative microorganisms from soil and the submerged fermentation study[J]. Journal of Fisheries of China,2016,40(10):1634-1642.
[16]李振高,骆永明,腾应.土壤与环境微生物研究法[M].北京:科学出版社,2008:52-61.Li Z G, Luo Y M, Teng Y. Soil and environmental microbiological research method[M]. Beijing:Science Press, 2008:52-61.
[17] GB11220.1-1989土壤中铀的测定CL-5209萃淋树脂分离2-(5-溴-2-吡啶偶氮)-5-二乙氨基苯酚分光光度法[S].GB 11220.1-1989 Determination of uranium in soil CL-5209extractant-containing resin separation 2-(5-bromo-2-pyridulazo)-5-diethlaminopheol spectrophotometry[S].
[18] Heinonen J K,Lahti R J. A new and convenient colorimetric determination of inorganic orthophosphate and its application to the assay of inorganic pyrophosphatase[J]. Analytical biochemistry, 1981,113(2):313-317.
[19]齐祖同.中国真菌志(第5卷):曲霉属及其相关有性型[M].北京:科学出版社,1997:92-93.Qi Z T. Chinese Fungi(5):Aspergillus and related sexual type[M].Beijing:Science Press,1997:92-93.
[20]吴沣,郝瑞霞,鲁安怀,等.塔宾曲霉的生物学特征及其对环境中Pb2+的固定作用研究[J].环境科学学报,2015,35(1):144-151.Wu F, Hao R X, Lu A H, et al. Biological characteristics of Aspergillus tubingensis and its fixation to Pb2+[J]. Acta Scientiae Circumstantiae,2015,35(1):144-151.
[21]刘晓芳,黄晓东,张芳.一株溶磷黑曲霉的溶磷特性及溶磷机制初探[J].河南农业科学,2005,34(6):60-62.Liu X F, Hang X D, Zhang F. Preliminary study on phosphatesolubilizing characteristic and mechanism of an Aspergillus niger strain[J]. Journal of Henan Agricultural Sciences, 2005,34(6):60-62.
[22] Rodriguez H, Fraga R. Phosphate solubilizing bacteria and their role in plant growth promotion[J]. Biotechnology advances, 1999,17(4/5):319-339.
[23] Illmer P, Schinner F. Solubilization of inorganic phosphates by microorganisms isolated from forest soils[J]. Soil Biology&Biochemistry, 1992,24(4):389-395.
[24]扬慧,范丙全,龚明波,等.一株新的溶磷草生欧文氏菌的分离、鉴定及其溶磷效果的初步研究[J].微生物学报,2008,48(1):51-56.Yang H, Fan B Q, Gong M B, et al. Isolation and identification of a novel phosphate-dissolving strain P21[J]. Acta Microbiologica Sinica,2008,48(1):51-56.
[25] Seaman J C, Hutchison J M, Jackson B P, et al. In situ treatment of metals in contaminated soils with phytate[J]. Journal of Environmental Quality, 2003,32(1):153-61.
[26]司慧,罗学刚,望子龙,等.枯草芽孢杆菌对铀的富集及机理研究[J].中国农学通报,2017,33(8):31-38.Si H, Luo X G, Wang Z L, et al. Biosorption of uranium by Bacillus subtil is and its mechanism[J]. Chinese Agricultural Science Bulletin,2017,33(8):31-38.
[27] Bachmaf S, Planer-Friedrich B, Merkel B J. Effect of sulfate,carbonate, and phosphate on the uranium(Ⅵ)sorption behavior onto bentonite[J]. Radiochimica Acta,2008,96(6):359-366.
[28] Yuan F, Wu C, Cai Y, et al. Synthesis of phytic acid-decorated titanate nanotubes for high efficient and high selective removal of U(Ⅵ)[J]. Chemical Engineering Journal, 2017,322:353-365.
[29] Hu N, Li K, Sui Y,et al. Utilization of phosphate rock as a sole source of phosphorus for uranium biomineralization mediated by Penicillium funiculosum[J]. RSC Advances, 2018,8(24):13459-13465.
[30] Ding D X, Xin X, Li L, et al. Removal and recovery of U(Ⅵ)from low concentration radioactive wastewater by ethylenediaminemodified biomass of Aspergillus niger[J]. Water, Air,&Soil Pollution, 2014,225(12):1-16.
[31] Choudhary S, Sar P. Uranium biomineralization by a metal resistant Pseudomonas aeruginosa strain isolated from contaminated mine waste[J]. Journal of hazardous materials, 2011,186(1):336-343.
[32] Zhang J, Song H,Chen Z,et al. Biomineralization mechanism of U(VI)induced by Bacillus cereus 12-2:The role of functional groups and enzymes[J]. Chemosphere, 2018,206:682-692.
[33] Zheng X Y, Shen Y H, Wang X Y, et al. Effect of pH on uranium(VI)biosorption and biomineralization by Saccharomyces cerevisiae[J].Chemosphere, 2018,203:109-116.
[34] Latta D E,Kemner K M,Mishra B,et al. Effects of calcium and phosphate on uranium(IV)oxidation:comparison between nanoparticulate uraninite and amorphous U IV-phosphate[J].Geochimica Et Cosmochimica Acta, 2016,174:122-142.
[2] Yuan F, Wu C, Cai Y, et al. Synthesis of phytic acid-decorated titanate nanotubes for high efficient and high selective removal ofU(VI)[J]. Chemical Engineering Journal, 2017,322:353-365
[3] Parrish R R, Horstwood M, Arnason J G, et al. Depleted uranium contamination by inhalation exposure and its detection after~20 years:implications for human health assessment[J]. Science of the Total Environment, 2008,390(1):58-68.
[4] Taylor D M, Taylor S K. Environmental uranium and human health[J].Reviews on Environmental Health, 1997,12(3):147-158.
[5] Newsome L,Morris K, Lloyd J R. The biogeochemistry and bioremediation of uranium and other priority radionuclides[J].Chemical Geology, 2014,363:164-184.
[6] Beazley M J, Martinez R J, Sobecky P A, et al. Uranium biomineralization as a result of bacterial phosphatase activity:insights from bacterial isolates from a contaminated subsurface[J].Environmental Science&Technology, 2007,41(16):5701-5707.
[7] Wu W, Carley J, Luo J, et al. In situ bioreduction of uranium(VI)to submicromolar levels and reoxidation by dissolved oxygen[J].Environmental Science&Technology, 2007,41(16):5716-5723.
[8] Moon H S, Komlos J, Jaffe P R. Biogenic U(IV)oxidation by dissolved oxygen and nitrate in sediment after prolonged U(VI)/Fe(Ⅲ)/SO42-reduction[J]. Journal of Contaminant Hydrology, 2009,105(1):18-27.
[9] Wan J, Tokunaga T K, Brodie E, et al. Reoxidation of bioreduced uranium under reducing conditions[J]. Environmental Science&Technology,2005,39(16):6162-6169.
[10] Wu W, Carley J, Fienen M, et al. Pilot-scale in situ bioremediation of uranium in a highly contaminated aquifer. 1. conditioning of a treatment zone[J]. Environmental Science&Technology, 2006,40(12):3978-3985.
[11] Wufuer R, Wei Y, Lin Q,et al. Uranium bioreduction and biomineralization[J]. Advances in Applied Microbiology, 2017,101:137-168.
[12] Knox A S,Brigmon R L, Kaplan D I, et al. Interactions among phosphate amendments, microbes and uranium mobility in contaminated sediments[J]. Science of the Total Environment,2008,395(2):63-71.
[13] Liang X,Hillier S, Pendlowski H, et al. Uranium phosphate biomineralization by fungi[J]. Environmental Microbiology, 2015,17(6):2064-2075.
[14] Salome K. Non-reductive biomineralization of U(Ⅵ)-phosphate minerals through the activities of microbial phytases[D]. Atlanta:Georgia Institute of Technology, 2013:48-52.
[15]王晓东,胡海滨,麦康森,等.土壤中单宁和植酸降解菌的筛选、鉴定及液态发酵研究[J].水产学报,2016,40(10):1634-1642.Wang X D, Hu H B, Mai K S, et al. Screening and identification of tannic acid and phytic acid degradative microorganisms from soil and the submerged fermentation study[J]. Journal of Fisheries of China,2016,40(10):1634-1642.
[16]李振高,骆永明,腾应.土壤与环境微生物研究法[M].北京:科学出版社,2008:52-61.Li Z G, Luo Y M, Teng Y. Soil and environmental microbiological research method[M]. Beijing:Science Press, 2008:52-61.
[17] GB11220.1-1989土壤中铀的测定CL-5209萃淋树脂分离2-(5-溴-2-吡啶偶氮)-5-二乙氨基苯酚分光光度法[S].GB 11220.1-1989 Determination of uranium in soil CL-5209extractant-containing resin separation 2-(5-bromo-2-pyridulazo)-5-diethlaminopheol spectrophotometry[S].
[18] Heinonen J K,Lahti R J. A new and convenient colorimetric determination of inorganic orthophosphate and its application to the assay of inorganic pyrophosphatase[J]. Analytical biochemistry, 1981,113(2):313-317.
[19]齐祖同.中国真菌志(第5卷):曲霉属及其相关有性型[M].北京:科学出版社,1997:92-93.Qi Z T. Chinese Fungi(5):Aspergillus and related sexual type[M].Beijing:Science Press,1997:92-93.
[20]吴沣,郝瑞霞,鲁安怀,等.塔宾曲霉的生物学特征及其对环境中Pb2+的固定作用研究[J].环境科学学报,2015,35(1):144-151.Wu F, Hao R X, Lu A H, et al. Biological characteristics of Aspergillus tubingensis and its fixation to Pb2+[J]. Acta Scientiae Circumstantiae,2015,35(1):144-151.
[21]刘晓芳,黄晓东,张芳.一株溶磷黑曲霉的溶磷特性及溶磷机制初探[J].河南农业科学,2005,34(6):60-62.Liu X F, Hang X D, Zhang F. Preliminary study on phosphatesolubilizing characteristic and mechanism of an Aspergillus niger strain[J]. Journal of Henan Agricultural Sciences, 2005,34(6):60-62.
[22] Rodriguez H, Fraga R. Phosphate solubilizing bacteria and their role in plant growth promotion[J]. Biotechnology advances, 1999,17(4/5):319-339.
[23] Illmer P, Schinner F. Solubilization of inorganic phosphates by microorganisms isolated from forest soils[J]. Soil Biology&Biochemistry, 1992,24(4):389-395.
[24]扬慧,范丙全,龚明波,等.一株新的溶磷草生欧文氏菌的分离、鉴定及其溶磷效果的初步研究[J].微生物学报,2008,48(1):51-56.Yang H, Fan B Q, Gong M B, et al. Isolation and identification of a novel phosphate-dissolving strain P21[J]. Acta Microbiologica Sinica,2008,48(1):51-56.
[25] Seaman J C, Hutchison J M, Jackson B P, et al. In situ treatment of metals in contaminated soils with phytate[J]. Journal of Environmental Quality, 2003,32(1):153-61.
[26]司慧,罗学刚,望子龙,等.枯草芽孢杆菌对铀的富集及机理研究[J].中国农学通报,2017,33(8):31-38.Si H, Luo X G, Wang Z L, et al. Biosorption of uranium by Bacillus subtil is and its mechanism[J]. Chinese Agricultural Science Bulletin,2017,33(8):31-38.
[27] Bachmaf S, Planer-Friedrich B, Merkel B J. Effect of sulfate,carbonate, and phosphate on the uranium(Ⅵ)sorption behavior onto bentonite[J]. Radiochimica Acta,2008,96(6):359-366.
[28] Yuan F, Wu C, Cai Y, et al. Synthesis of phytic acid-decorated titanate nanotubes for high efficient and high selective removal of U(Ⅵ)[J]. Chemical Engineering Journal, 2017,322:353-365.
[29] Hu N, Li K, Sui Y,et al. Utilization of phosphate rock as a sole source of phosphorus for uranium biomineralization mediated by Penicillium funiculosum[J]. RSC Advances, 2018,8(24):13459-13465.
[30] Ding D X, Xin X, Li L, et al. Removal and recovery of U(Ⅵ)from low concentration radioactive wastewater by ethylenediaminemodified biomass of Aspergillus niger[J]. Water, Air,&Soil Pollution, 2014,225(12):1-16.
[31] Choudhary S, Sar P. Uranium biomineralization by a metal resistant Pseudomonas aeruginosa strain isolated from contaminated mine waste[J]. Journal of hazardous materials, 2011,186(1):336-343.
[32] Zhang J, Song H,Chen Z,et al. Biomineralization mechanism of U(VI)induced by Bacillus cereus 12-2:The role of functional groups and enzymes[J]. Chemosphere, 2018,206:682-692.
[33] Zheng X Y, Shen Y H, Wang X Y, et al. Effect of pH on uranium(VI)biosorption and biomineralization by Saccharomyces cerevisiae[J].Chemosphere, 2018,203:109-116.
[34] Latta D E,Kemner K M,Mishra B,et al. Effects of calcium and phosphate on uranium(IV)oxidation:comparison between nanoparticulate uraninite and amorphous U IV-phosphate[J].Geochimica Et Cosmochimica Acta, 2016,174:122-142.