Two selenium tolerant Lysinibacillus sp. strains are capable of reducing selenite to elemental Se efficiently under aerobic conditions
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摘要
Microbes play important roles in the transport and transformation of selenium(Se) in the environment, thereby influencing plant resistance to Se and Se accumulation in plant. The objectives are to characterize the bacteria with high Se tolerance and reduction capacity and explore the significance of microbial origins on their Se tolerance, reduction rate and efficiency. Two bacterial strains were isolated from a naturally occurred Se-rich soil at tea orchard in southern Anhui Province, China. The reduction kinetics of selenite was investigated and the reducing product was characterized using scanning electron microscopy and transmission electron microscopy-energy dispersive spectroscopy. The bacteria were identified as Lysinibacillus xylanilyticus and Lysinibacillus macrolides,respectively, using morphological, physiological and molecular methods. The results showed that the minimal inhibitory concentrations(MICs) of selenite for L. xylanilyticus and L. macrolides were 120 and 220 mmol/L, respectively, while MICs of selenate for L.xylanilyticus and L. macrolides were 800 and 700 mmol/L, respectively. Both strains aerobically reduced selenite with an initial concentration of 1.0 mmol/L to elemental Se nanoparticles(SeNPs) completely within 36 hr. Biogenic SeNPs were observed both inside and outside the cells suggesting either an intra-or extracellular reduction process. Our study implied that the microbes from Se-rich environments were more tolerant to Se and generally quicker and more efficient than those from Se-free habitats in the reduction of Se oxyanions. The bacterial strains with high Se reduction capacity and the biological synthesized Se NPs would have potential applications in agriculture, food, environment and medicine.
Microbes play important roles in the transport and transformation of selenium(Se) in the environment, thereby influencing plant resistance to Se and Se accumulation in plant. The objectives are to characterize the bacteria with high Se tolerance and reduction capacity and explore the significance of microbial origins on their Se tolerance, reduction rate and efficiency. Two bacterial strains were isolated from a naturally occurred Se-rich soil at tea orchard in southern Anhui Province, China. The reduction kinetics of selenite was investigated and the reducing product was characterized using scanning electron microscopy and transmission electron microscopy-energy dispersive spectroscopy. The bacteria were identified as Lysinibacillus xylanilyticus and Lysinibacillus macrolides,respectively, using morphological, physiological and molecular methods. The results showed that the minimal inhibitory concentrations(MICs) of selenite for L. xylanilyticus and L. macrolides were 120 and 220 mmol/L, respectively, while MICs of selenate for L.xylanilyticus and L. macrolides were 800 and 700 mmol/L, respectively. Both strains aerobically reduced selenite with an initial concentration of 1.0 mmol/L to elemental Se nanoparticles(SeNPs) completely within 36 hr. Biogenic SeNPs were observed both inside and outside the cells suggesting either an intra-or extracellular reduction process. Our study implied that the microbes from Se-rich environments were more tolerant to Se and generally quicker and more efficient than those from Se-free habitats in the reduction of Se oxyanions. The bacterial strains with high Se reduction capacity and the biological synthesized Se NPs would have potential applications in agriculture, food, environment and medicine.
Microbes play important roles in the transport and transformation of selenium(Se) in the environment, thereby influencing plant resistance to Se and Se accumulation in plant. The objectives are to characterize the bacteria with high Se tolerance and reduction capacity and explore the significance of microbial origins on their Se tolerance, reduction rate and efficiency. Two bacterial strains were isolated from a naturally occurred Se-rich soil at tea orchard in southern Anhui Province, China. The reduction kinetics of selenite was investigated and the reducing product was characterized using scanning electron microscopy and transmission electron microscopy-energy dispersive spectroscopy. The bacteria were identified as Lysinibacillus xylanilyticus and Lysinibacillus macrolides,respectively, using morphological, physiological and molecular methods. The results showed that the minimal inhibitory concentrations(MICs) of selenite for L. xylanilyticus and L. macrolides were 120 and 220 mmol/L, respectively, while MICs of selenate for L.xylanilyticus and L. macrolides were 800 and 700 mmol/L, respectively. Both strains aerobically reduced selenite with an initial concentration of 1.0 mmol/L to elemental Se nanoparticles(SeNPs) completely within 36 hr. Biogenic SeNPs were observed both inside and outside the cells suggesting either an intra-or extracellular reduction process. Our study implied that the microbes from Se-rich environments were more tolerant to Se and generally quicker and more efficient than those from Se-free habitats in the reduction of Se oxyanions. The bacterial strains with high Se reduction capacity and the biological synthesized Se NPs would have potential applications in agriculture, food, environment and medicine.
引文
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Avenda?o,R.,Chaves,N.,Fuentes,P.,Sánchez,E.,Jiménez,J.I.,Chavarría,M.,2016.Production of selenium nanoparticles in Pseudomonas putida KT2440.Sci.Rep.6,37155.
Ayano,H.,Miyake,M.,Terasawa,K.,Kuroda,M.,Soda,S.,Sakaguchi,T.,et al.,2014.Isolation of a selenite-reducing and cadmium-resistant bacterium Pseudomonas sp.strain RB for microbial synthesis of CdSe nanoparticles.J.Biosci.Bioeng.117(5),576-581.
Bao,P.,Xiao,K.Q.,Wang,H.J.,Xu,H.,Xu,P.P.,Jia,Y.,et al.,2016.Characterization and potential applications of a selenium nanoparticle producing and nitrate reducing bacterium Baccilus oryziterrae sp.nov.Sci.Rep.6,34054.
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Compilation Committee of the Atlas of Endemic Diseases and Their Environments in the People's Republic of China,1989.The atlas of endemic diseases and their environments in the People's Republic of China.Science Press,Beijing.
Coorevits,A.,Dinsdale,A.E.,Heyrman,J.,Schumann,P.,Van Landschoot,A.,Logan,N.A.,et al.,2012.Lysinibaccillus macroides sp nov.,nom.rev.Int.J.Syst.Evol.Microbiol.62,1121-1127.
Debieux,C.M.,Dridge,E.J.,Mueller,C.M.,Splatt,P.,Paszkiewicz,K.,Knight,I.,et al.,2011.A bacterial process for selenium nanosphere assembly.Proc.Natl.Acad.Sci.U.S.A.108(33),13480-13485.
Dhanjal,S.,Cameotra,S.S.,2010.Aerobic biogenesis of selenium nanospheres by Bacillus cereus isolated from coalmine soil.Microb.Cell Factories 9(1),52.
Ding,Y.Z.,Wang,R.G.,Guo,J.K.,Wu,F.C.,Xu,Y.M.,Feng,R.W.,2015.The effect of selenium on the subcellular distribution of antimony to regulate the toxicity of antimony in paddy soil.Environ.Sci.Pollut.Res.22(7),5111-5123.
Durán,P.,Acu?a,J.J.,Gianfreda,L.,Azcón,R.,Funes-Collado,V.,Mora,M.L.,2015.Endophytic selenobacteria as new inocula for selenium biofortification.Appl.Soil Ecol.96,319-326.
Dwivedi,S.,Alkhedhairy,A.A.,Ahamed,M.,Musarrat,J.,2013.Biomimetic synthesis of selenium nanospheres by bacterial strain JS-11 and its role as a biosensor for nanotoxicity assessment:a novel Se-bioassay.PLoS One 8,e57404.
Espinosa-Ortiz,E.J.,Shakya,M.,Jain,R.,Rene,E.R.,van Hullebusch,E.D.,Lens,P.N.L.,2016.Sorption of zinc onto elemental selenium nanoparticles immobilized in Phanerochaete chrysosporiumpellets.Environ.Sci.Pollut.Res.23(21),21619-21630.
Gao,J.,Liu,Y.,Huang,Y.,Lin,Z.Q.,Ba?uelos,G.S.,Lam,M.H.W.,et al.,2011.Daily selenium intake in a moderate selenium deficiency area of Suzhou,China.Food Chem.126(3),1088-1093.
Ghosh,A.,Mohod,A.M.,Paknikar,K.M.,Jain,R.K.,2008.Isolation and characterization of selenite-and selenate-tolerant microorganisms from selenium-contaminated sites.World J.Microbiol.Biotechnol.24(6),1607-1611.
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Huang,X.Q.,Chen,X.,Chen,Q.C.,Yu,Q.Q.,Sun,D.D.,Liu,J.,2016.Investigation of functional selenium nanoparticles as potent antimicrobial agents against superbugs.Acta Biomater.30,397-407.
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Jiang,H.D.,He,X.H.,Zhang,L.X.,Tao,Y.,Wang,X.M.,Gao,P.,et al.,2010.Reduction of selenite to elemental red selenium under aerobic condition by Pseudomonas alcaliphila MBR.Acta Microbiol Sin.50(10),1347-1352.
Kashiwa,M.,Nishimoto,S.,Takahashi,K.,Ike,M.,Fujita,M.,2000.Factors affecting soluble selenium removal by a selenate-reducing bacterium Bacillus sp.SF-1.J.Biosci.Bioeng.89(6),528-533.
Kessi,J.,2006.Enzymic systems proposed to be involved in the dissimilatory reduction of selenite in the purple non-sulfur bacteria Rhodospirillum rubrum and Rhodobacter capsulatus.Microbiol.Soc.J.152,731-743.
Kessi,J.,Hanselmann,K.W.,2004.Similarities between the abiotic reduction of selenite with glutathione and the dissimilatory reaction mediated by Rhodospirillum rubrum and Escherichia coli.J.Biol.Chem.279(49),50662-50669.
Khoei,N.S.,Lampis,S.,Zonaro,E.,Yrj?l?,K.,Bernardi,P.,Vallini,G.,2017.Insights into selenite reduction and biogenesis of elemental selenium nanoparticles by two environmental isolates of Burkholderia fungorum.New Biotechnol.34,1-11.
Kumar,S.,Nei,M.,Dudley,J.,Tamura,K.,2008.MEGA:a biologistcentric software for evolutionary analysis of DNA and protein sequences.Brief.Bioinform.9(4),299-306.
Kumar,A.,Singh,R.P.,Singh,P.K.,Awasthi,S.,Chakrabarty,D.,Trivedi,P.K.,et al.,2014.Selenium ameliorates arsenic induced oxidative stress through modulation of antioxidant enzymes and thiols in rice(Oryza sativa L.).Ecotoxicology 23(7),1153-1163.
Lampis,S.,Zonaro,E.,Bertolini,C.,Bernardi,P.,Butler,C.S.,Vallini,G.,2014.Delayed formation of zero-valent selenium nanoparticles by Bacillus mycoides SeITE01 as a consequence of selenite reduction under aerobic conditions.Microb.Cell Factories 13,35.
Lampis,S.,Zonaro,E.,Bertolini,C.,Cecconi,D.,Monti,F.,Micaroni,M.,et al.,2017.Selenite biotransformation and detoxification by Stenotrophomonas maltophilia SeITE02:novel clues on the route to bacterial biogenesis of selenium nanoparticles.J.Hazard.Mater.324,3-14.
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Bao,P.,Xiao,K.Q.,Wang,H.J.,Xu,H.,Xu,P.P.,Jia,Y.,et al.,2016.Characterization and potential applications of a selenium nanoparticle producing and nitrate reducing bacterium Baccilus oryziterrae sp.nov.Sci.Rep.6,34054.
Bettinelli,M.,Beone,G.M.,Spezia,S.,Baffi,C.,2000.Determination of heavy metals in soils and sediments by microwave-assisted digestion and inductively coupled plasma optical emission spectrometry analysis.Anal.Chim.Acta 424(2),289-296.
Combs,G.F.,2001.Selenium in global food systems.Br.J.Nutr.85(5),517-547.
Compilation Committee of the Atlas of Endemic Diseases and Their Environments in the People's Republic of China,1989.The atlas of endemic diseases and their environments in the People's Republic of China.Science Press,Beijing.
Coorevits,A.,Dinsdale,A.E.,Heyrman,J.,Schumann,P.,Van Landschoot,A.,Logan,N.A.,et al.,2012.Lysinibaccillus macroides sp nov.,nom.rev.Int.J.Syst.Evol.Microbiol.62,1121-1127.
Debieux,C.M.,Dridge,E.J.,Mueller,C.M.,Splatt,P.,Paszkiewicz,K.,Knight,I.,et al.,2011.A bacterial process for selenium nanosphere assembly.Proc.Natl.Acad.Sci.U.S.A.108(33),13480-13485.
Dhanjal,S.,Cameotra,S.S.,2010.Aerobic biogenesis of selenium nanospheres by Bacillus cereus isolated from coalmine soil.Microb.Cell Factories 9(1),52.
Ding,Y.Z.,Wang,R.G.,Guo,J.K.,Wu,F.C.,Xu,Y.M.,Feng,R.W.,2015.The effect of selenium on the subcellular distribution of antimony to regulate the toxicity of antimony in paddy soil.Environ.Sci.Pollut.Res.22(7),5111-5123.
Durán,P.,Acu?a,J.J.,Gianfreda,L.,Azcón,R.,Funes-Collado,V.,Mora,M.L.,2015.Endophytic selenobacteria as new inocula for selenium biofortification.Appl.Soil Ecol.96,319-326.
Dwivedi,S.,Alkhedhairy,A.A.,Ahamed,M.,Musarrat,J.,2013.Biomimetic synthesis of selenium nanospheres by bacterial strain JS-11 and its role as a biosensor for nanotoxicity assessment:a novel Se-bioassay.PLoS One 8,e57404.
Espinosa-Ortiz,E.J.,Shakya,M.,Jain,R.,Rene,E.R.,van Hullebusch,E.D.,Lens,P.N.L.,2016.Sorption of zinc onto elemental selenium nanoparticles immobilized in Phanerochaete chrysosporiumpellets.Environ.Sci.Pollut.Res.23(21),21619-21630.
Gao,J.,Liu,Y.,Huang,Y.,Lin,Z.Q.,Ba?uelos,G.S.,Lam,M.H.W.,et al.,2011.Daily selenium intake in a moderate selenium deficiency area of Suzhou,China.Food Chem.126(3),1088-1093.
Ghosh,A.,Mohod,A.M.,Paknikar,K.M.,Jain,R.K.,2008.Isolation and characterization of selenite-and selenate-tolerant microorganisms from selenium-contaminated sites.World J.Microbiol.Biotechnol.24(6),1607-1611.
Güven,K.,Mutlu,M.B.,?irpan,C.,Kutlu,H.M.,2013.Isolation and identification of selenite reducing archaea from Tuz(salt)lake in Turkey.J.Basic Microbiol.53(5),397-401.
Huang,X.Q.,Chen,X.,Chen,Q.C.,Yu,Q.Q.,Sun,D.D.,Liu,J.,2016.Investigation of functional selenium nanoparticles as potent antimicrobial agents against superbugs.Acta Biomater.30,397-407.
Ike,M.,Takahashi,K.,Fujita,T.,Kashiwa,M.,Fujita,M.,2000.Selenate reduction by bacteria isolated from aquatic environment free from selenium contamination.Water Res.34(11),3019-3025.
IUSS Working Group WRB,2015.World Reference Base for soil resources 2014.Update.2015 International soil classification system for naming soils and creating legends for soil maps.World soil resources reports no.106.FAO,Rome.
Jiang,H.D.,He,X.H.,Zhang,L.X.,Tao,Y.,Wang,X.M.,Gao,P.,et al.,2010.Reduction of selenite to elemental red selenium under aerobic condition by Pseudomonas alcaliphila MBR.Acta Microbiol Sin.50(10),1347-1352.
Kashiwa,M.,Nishimoto,S.,Takahashi,K.,Ike,M.,Fujita,M.,2000.Factors affecting soluble selenium removal by a selenate-reducing bacterium Bacillus sp.SF-1.J.Biosci.Bioeng.89(6),528-533.
Kessi,J.,2006.Enzymic systems proposed to be involved in the dissimilatory reduction of selenite in the purple non-sulfur bacteria Rhodospirillum rubrum and Rhodobacter capsulatus.Microbiol.Soc.J.152,731-743.
Kessi,J.,Hanselmann,K.W.,2004.Similarities between the abiotic reduction of selenite with glutathione and the dissimilatory reaction mediated by Rhodospirillum rubrum and Escherichia coli.J.Biol.Chem.279(49),50662-50669.
Khoei,N.S.,Lampis,S.,Zonaro,E.,Yrj?l?,K.,Bernardi,P.,Vallini,G.,2017.Insights into selenite reduction and biogenesis of elemental selenium nanoparticles by two environmental isolates of Burkholderia fungorum.New Biotechnol.34,1-11.
Kumar,S.,Nei,M.,Dudley,J.,Tamura,K.,2008.MEGA:a biologistcentric software for evolutionary analysis of DNA and protein sequences.Brief.Bioinform.9(4),299-306.
Kumar,A.,Singh,R.P.,Singh,P.K.,Awasthi,S.,Chakrabarty,D.,Trivedi,P.K.,et al.,2014.Selenium ameliorates arsenic induced oxidative stress through modulation of antioxidant enzymes and thiols in rice(Oryza sativa L.).Ecotoxicology 23(7),1153-1163.
Lampis,S.,Zonaro,E.,Bertolini,C.,Bernardi,P.,Butler,C.S.,Vallini,G.,2014.Delayed formation of zero-valent selenium nanoparticles by Bacillus mycoides SeITE01 as a consequence of selenite reduction under aerobic conditions.Microb.Cell Factories 13,35.
Lampis,S.,Zonaro,E.,Bertolini,C.,Cecconi,D.,Monti,F.,Micaroni,M.,et al.,2017.Selenite biotransformation and detoxification by Stenotrophomonas maltophilia SeITE02:novel clues on the route to bacterial biogenesis of selenium nanoparticles.J.Hazard.Mater.324,3-14.
Lane,D.J.,1991.16S/23S rRNA sequencing.In:Stackebrandt,E.,Goodfellow,M.(Eds.),Nucleic acid techniques in bacterial systematics.John Wiley and Sons,New York,pp.115-175.
Levander,O.A.,Burk,R.F.,2006.Uptake of human dietary standards for selenium.In:Hatfield,D.L.,Berry,M.J.,Gladyshev,V.N.(Eds.),Selenium:its molecular biology and role in human health,2nd ed.Springer,New York,pp.399-410.
Li,B.Z.,Liu,N.,Li,Y.Q.,Jing,W.X.,Fan,J.H.,Li,D.,et al.,2014.Reduction of selenite to red elemental selenium by Rhodopseudomonas palustris strain N.PLoS One 9,e95955.
Lindblom,S.D.,Fakra,S.C.,Landon,J.,Schulz,P.,Tracy,B.,PilonSmits,E.A.H.,2014.Inoculation of selenium hyperaccumulator Stanleya pinnata and related non-accumulator Stanleya elata with hyperaccumulator rhizosphere fungi-investigation of effects on Se accumulation and speciation.Physiol.Plant.150(1),107-118.
Lu,R.K.,2000.Analysis method on soil agricultural chemistry.China Agricultural Science And Technology Press,Beijing.
Mal,J.,Veneman,W.J.,Nancharaiah,Y.V.,van Hullebusch,E.D.,Peijnenburg,W.J.G.M.,Vijver,M.G.,et al.,2017.A comparison of fate and toxicity of selenite,biogenically,and chemically synthesized selenium nanoparticles to zebrafish(Danio rerio)embryogenesis.Nanotoxicology 11(1),87-97.
Nagy,G.,Benko,I.,Kiraly,G.,Voros,O.,Tanczos,B.,Sztrik,A.,et al.,2015.Cellular and nephrotoxicity of selenium species.J.Trace Elem.Med.Biol.30,160-170.
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