一株耐盐硫氧化细菌的分离鉴定及脱硫机理
|
摘要
筛选高效脱硫菌株并揭示其去除硫化物的机理对生物脱硫技术的发展具有重要意义.从天然气净化厂污水站废水中分离出一株化能自养型硫氧化细菌,命名为DS-B.根据16S r DNA基因序列相似性分析,结合形态特征,对该菌株进行鉴定.进一步研究DS-B的硫代硫酸盐耐受性,并通过测定培养体系中各价态含硫物质的含量,探讨该菌的硫氧化性能.结果显示,菌株DS-B与Thioalkalivibrio thiocyanoxidans ARh2的序列相似性最高,初步鉴定其属于Thioalkalivibrio.DS-B的最佳生长和降解温度为35℃,35℃下培养7 d,硫代硫酸盐(S2O32-)的去除率能达到98.7%.该菌对S2O32-有较强的耐受性,且最适S2O32-浓度为2×104 mg/L.此外,对各价态硫氧化物的分析表明该菌氧化S2O32-的途径为:S2O32-→SO42-/S2O32-→S→SO42-.上述表明,菌株DS-B具有较强的耐盐能力,对硫代硫酸盐具有较高的去除能力,是一株具有应用价值的硫氧化细菌.
Micro-organism with efficient desulfurization performance is a key factor in the biological desulfurization technology.This study aimed to seek such a sulfur-oxidizing strain and understand its desulfurization mechanism.Wastewater in a sewage station of natural gas purification plant was used to screen the sulfide-oxidizing strain,and to identify it based on sequence similarity analysis of 16 S rDNA and the morphological characteristics.Thiosulfate was used as substrate for investigating the sulfur oxidation performance and salinity tolerance;the OD_(600),content change of thiosulfate,sulfate,sulfur,pH and total alkalinity in the cultural system were also investigated.The strain DS-B was found to share the highest sequence similarity with Thioalkalivibrio thiocyanoxidans ARh2,therefore determined as Thioalkalivibrio.At the optimum temperature of 35 ℃ for growth and degradation,the removal efficiency of thiosulfate could reach 98.7%after 7 days.Strain DS-B had strong resistance to thiosulfate,and the optimal concentration of S_2O_3~(2-) was 2 x 10~4 mg/L.The analysis for sulfur oxides showed that it could oxidize thiosulfate by the pathway of S_2O_3~(2-)→SO_4~(2-) / S_2O_3~(2-)→S→SO_4~(2-).Therefore the strain DS-B is a sulfur-oxidizing bacterium with great application prospect for its strong salt tolerance and conspicuous removal capability for thiosulfate.
Micro-organism with efficient desulfurization performance is a key factor in the biological desulfurization technology.This study aimed to seek such a sulfur-oxidizing strain and understand its desulfurization mechanism.Wastewater in a sewage station of natural gas purification plant was used to screen the sulfide-oxidizing strain,and to identify it based on sequence similarity analysis of 16 S rDNA and the morphological characteristics.Thiosulfate was used as substrate for investigating the sulfur oxidation performance and salinity tolerance;the OD_(600),content change of thiosulfate,sulfate,sulfur,pH and total alkalinity in the cultural system were also investigated.The strain DS-B was found to share the highest sequence similarity with Thioalkalivibrio thiocyanoxidans ARh2,therefore determined as Thioalkalivibrio.At the optimum temperature of 35 ℃ for growth and degradation,the removal efficiency of thiosulfate could reach 98.7%after 7 days.Strain DS-B had strong resistance to thiosulfate,and the optimal concentration of S_2O_3~(2-) was 2 x 10~4 mg/L.The analysis for sulfur oxides showed that it could oxidize thiosulfate by the pathway of S_2O_3~(2-)→SO_4~(2-) / S_2O_3~(2-)→S→SO_4~(2-).Therefore the strain DS-B is a sulfur-oxidizing bacterium with great application prospect for its strong salt tolerance and conspicuous removal capability for thiosulfate.
引文
1王钢,王欣,高德玉,刘伟,赫大新,陈薇.沼气生物脱硫技术研究[J].应用能源技术,2005,33(5):33-35[Wang G,Wang X,Gao DY,Liu W,He DX,Chen W.Study on biological desulfurization technology for biogas[J].Appl Energy Technol,2005,33(5):33-35]
2李国欣,杨俊仕,李旭东.天然气净化厂的废水处理设计与运行[J].中国给水排水,2009,25(22):49-55[Li GX,Yang JS,Li XD.Design and operation of wastewater treatment process in natural gas purification plant[J].China Water Wastewater,2009,25(22):49-55]
3蒲万芬,胡佩.酸性气体中硫化氢的微生物脱除方法[J].天然气工业,2005,25(3):166-171[Pu WF,Hu P.Method to remove hydrogen sulfide from sour gas by microorganisms[J].Nat Gas Ind,2005,25(3):166-171]
4汪家铭.Shell-Paques生物脱硫技术及其应用[J].化肥设计,2010,48(2):39-42[Wang JM.Shell-Paques biodesulfur technology and its application[J].Chem Fertil Des,2010,48(2):39-42]
5李红菊,唐晓东,赵红义,郭巧霞.天然气脱硫技术[J].石油化工腐蚀与防护,2009,26(5):4-6[Li HJ,Tang XD,Zhao HY,Guo QX.Natural gas desulfurization technology[J].Corr Prot Petrochem Ind,2009,26(5):4-6]
6 Buisman CJN,Wit B,Lettinga G.Biotechnological sulfide removal in three polyurethane carrier reactors:strirred reactor,bio-rotor reactor and up-flow reactor[J].Water Res,1990,24(2):245-251
7李军幸,张克强,季民,袁志华,分光光度法测定微生物处理合硫化物废水中的单质硫含量[J].农业环境科学学报,2006,25(1):261-265[Li JX,Zhang KQ,Ji M,Yuan ZH.Spectrophotometric quantification of sulfur in sulfide-containing wastewater treated with microbial[J].J Agro Environ Sci,2006,25(1):261-265]
8王蕙祥,姜理英,吴晓薇,陈建孟.硫氧化细菌的分离鉴定及降解特性[J].应用与环境工程学报,2011,17(5):706-710[Wang HX,Jiang LY,Wu XW,Chen JM.Isolation,identification and degradation characteristics of a sulfide-oxidizing bacterium[J].Chin J Appl Environ Biol,2011,17(5):706-710]
9邓良伟,唐一,吴彦.生物脱硫机理及其研究进展[J].上海环境科学,1998,17(5):35-39[Deng LW,Tang Y,Wu Y.Mechanism of biological desulfurization and its progress[J].Shanghai Environ Sci,1998,17(5):35-39]
10顾运,杨宏,吴宣,尚海源.盐分对一株硫氧化细菌氧化能力的影响[J].环境工程学报,2015,9(1):491-494[Gu Y,Yang H,Wu X,Shang HY Effect of salinity on a sulfur-oxidizing bacteria oxidation ability[J].Chin J Environ Eng,2015,9(1):491-494]
11蒋永荣,胡明成.硫酸盐废水生物脱硫研究进展[J].环境科学与技术,2008,31(9):67-71[Jiang YR,Hu MC.Biological desulfurization of sulfate containing wastewater[J].Environ Sci Technol,31(9):67-71]
12王爱杰,杜大仲,任南琪,赵秋实,王文静.脱氮硫杆菌在废水脱硫、脱氮处理工艺中的应用[J].哈尔滨工业大学学报,2004,36(4):423-425,429[Wang AJ,Du DZ,Ren NQ,Zhao QS,Wang WJ.Application of Thiobacillus denitrificans in wastewater denitrification and de-sulfide treatment processes[J].J Harbin Inst Technol,2004,36(4):423-425,429]
13 Pethkar AV,Paknikar KM.Thiosulfate biodegradation/silver biosorption process for the treatment of photofilm processing wastewater.Process Biochem,2003,38(6):855-860
14 Lin BL,Hosomi M,Murakami A.Effects of high salinity and constitute organic compounds on treatment of photo-processing waste by a sulphur-oxidizing bacteria/granular activated carbon sludge system[J].Water Res,2002,36(4):1076-1083
15林栋青,张彦科,顾向阳.硫代硫酸盐氧化菌TX的分离、鉴定及其生物学特性[J].微生物学通报,2009,36(11):1638-1644[Lin DQ,Zhang YK,Gu XY.Isolation,identification and characterization of thiosulfateoxidizing bacterium TX[J].Microbiology,2009,36(11):1638-1644
16 Baalsrud K,Kjellrun S Baalsrud.Studies on Thiobacillus denitrificans[J].Archiv Microbiol,1954,20(1):34-62
17东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001:353-356[Dong XZ,Cai MY.System Identification Manual of Common Bacteria[M].Beijing:Science Press,2001:353-356]
18 Mahmood Q,Hu B,Cai J.Zheng P,Azim MR,Jilani G,Islam E.Isolation of Ochrobacte-rium sp.QZ2 from sulfide and nitrite treatment system[J].J Hazard Mater,2009,165(1-3):558-565
19国家环保局.水和废水监测分析方法[M].第4版,北京:中国环境科学出版社,2002[Ministry of Environmental Protection of China.Water and Wastewater Monitoring Analysis Method-IV[M].Beijing:Chinese Environmental Science Press,2002]
20李军幸,张克强,季民,袁志华.分光光度法测定微生物处理合硫化物废水中的单质硫合量[J].农业环境科学学报,2006,25(1):261-265[Li JX,Zhang KQ,Ji M,Yuan ZH.Spectrophotometric quantification of sulfur in sulfide-containing wastewater treated with microbial[J].J Agro Environ Sci,2006,25(1):261-265]
21沈镭,张再丽,贾晓珊.氧化亚铁硫杆菌和氧化硫硫杆菌对硫代硫酸钠的代谢机理研究[J].环境科学学报,2006,26(12):2000-2007[Shen L,Zhang ZL,Jia XS.Study on metabolic mechanisms of sodium thiosulfate by Thiobacillus Ferrooxidans and Thiobacillus Thiooxidans[J].J Environ Sci,2006,26(12):2000-2007]
22 Solicio C,Lodi A,Veglio F.Bioleaching of zinc and aluminium from industrial waste sludge by means of Thiobacillus ferrooxidans[J].Waste Mana,2002,22:667-675
2李国欣,杨俊仕,李旭东.天然气净化厂的废水处理设计与运行[J].中国给水排水,2009,25(22):49-55[Li GX,Yang JS,Li XD.Design and operation of wastewater treatment process in natural gas purification plant[J].China Water Wastewater,2009,25(22):49-55]
3蒲万芬,胡佩.酸性气体中硫化氢的微生物脱除方法[J].天然气工业,2005,25(3):166-171[Pu WF,Hu P.Method to remove hydrogen sulfide from sour gas by microorganisms[J].Nat Gas Ind,2005,25(3):166-171]
4汪家铭.Shell-Paques生物脱硫技术及其应用[J].化肥设计,2010,48(2):39-42[Wang JM.Shell-Paques biodesulfur technology and its application[J].Chem Fertil Des,2010,48(2):39-42]
5李红菊,唐晓东,赵红义,郭巧霞.天然气脱硫技术[J].石油化工腐蚀与防护,2009,26(5):4-6[Li HJ,Tang XD,Zhao HY,Guo QX.Natural gas desulfurization technology[J].Corr Prot Petrochem Ind,2009,26(5):4-6]
6 Buisman CJN,Wit B,Lettinga G.Biotechnological sulfide removal in three polyurethane carrier reactors:strirred reactor,bio-rotor reactor and up-flow reactor[J].Water Res,1990,24(2):245-251
7李军幸,张克强,季民,袁志华,分光光度法测定微生物处理合硫化物废水中的单质硫含量[J].农业环境科学学报,2006,25(1):261-265[Li JX,Zhang KQ,Ji M,Yuan ZH.Spectrophotometric quantification of sulfur in sulfide-containing wastewater treated with microbial[J].J Agro Environ Sci,2006,25(1):261-265]
8王蕙祥,姜理英,吴晓薇,陈建孟.硫氧化细菌的分离鉴定及降解特性[J].应用与环境工程学报,2011,17(5):706-710[Wang HX,Jiang LY,Wu XW,Chen JM.Isolation,identification and degradation characteristics of a sulfide-oxidizing bacterium[J].Chin J Appl Environ Biol,2011,17(5):706-710]
9邓良伟,唐一,吴彦.生物脱硫机理及其研究进展[J].上海环境科学,1998,17(5):35-39[Deng LW,Tang Y,Wu Y.Mechanism of biological desulfurization and its progress[J].Shanghai Environ Sci,1998,17(5):35-39]
10顾运,杨宏,吴宣,尚海源.盐分对一株硫氧化细菌氧化能力的影响[J].环境工程学报,2015,9(1):491-494[Gu Y,Yang H,Wu X,Shang HY Effect of salinity on a sulfur-oxidizing bacteria oxidation ability[J].Chin J Environ Eng,2015,9(1):491-494]
11蒋永荣,胡明成.硫酸盐废水生物脱硫研究进展[J].环境科学与技术,2008,31(9):67-71[Jiang YR,Hu MC.Biological desulfurization of sulfate containing wastewater[J].Environ Sci Technol,31(9):67-71]
12王爱杰,杜大仲,任南琪,赵秋实,王文静.脱氮硫杆菌在废水脱硫、脱氮处理工艺中的应用[J].哈尔滨工业大学学报,2004,36(4):423-425,429[Wang AJ,Du DZ,Ren NQ,Zhao QS,Wang WJ.Application of Thiobacillus denitrificans in wastewater denitrification and de-sulfide treatment processes[J].J Harbin Inst Technol,2004,36(4):423-425,429]
13 Pethkar AV,Paknikar KM.Thiosulfate biodegradation/silver biosorption process for the treatment of photofilm processing wastewater.Process Biochem,2003,38(6):855-860
14 Lin BL,Hosomi M,Murakami A.Effects of high salinity and constitute organic compounds on treatment of photo-processing waste by a sulphur-oxidizing bacteria/granular activated carbon sludge system[J].Water Res,2002,36(4):1076-1083
15林栋青,张彦科,顾向阳.硫代硫酸盐氧化菌TX的分离、鉴定及其生物学特性[J].微生物学通报,2009,36(11):1638-1644[Lin DQ,Zhang YK,Gu XY.Isolation,identification and characterization of thiosulfateoxidizing bacterium TX[J].Microbiology,2009,36(11):1638-1644
16 Baalsrud K,Kjellrun S Baalsrud.Studies on Thiobacillus denitrificans[J].Archiv Microbiol,1954,20(1):34-62
17东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001:353-356[Dong XZ,Cai MY.System Identification Manual of Common Bacteria[M].Beijing:Science Press,2001:353-356]
18 Mahmood Q,Hu B,Cai J.Zheng P,Azim MR,Jilani G,Islam E.Isolation of Ochrobacte-rium sp.QZ2 from sulfide and nitrite treatment system[J].J Hazard Mater,2009,165(1-3):558-565
19国家环保局.水和废水监测分析方法[M].第4版,北京:中国环境科学出版社,2002[Ministry of Environmental Protection of China.Water and Wastewater Monitoring Analysis Method-IV[M].Beijing:Chinese Environmental Science Press,2002]
20李军幸,张克强,季民,袁志华.分光光度法测定微生物处理合硫化物废水中的单质硫合量[J].农业环境科学学报,2006,25(1):261-265[Li JX,Zhang KQ,Ji M,Yuan ZH.Spectrophotometric quantification of sulfur in sulfide-containing wastewater treated with microbial[J].J Agro Environ Sci,2006,25(1):261-265]
21沈镭,张再丽,贾晓珊.氧化亚铁硫杆菌和氧化硫硫杆菌对硫代硫酸钠的代谢机理研究[J].环境科学学报,2006,26(12):2000-2007[Shen L,Zhang ZL,Jia XS.Study on metabolic mechanisms of sodium thiosulfate by Thiobacillus Ferrooxidans and Thiobacillus Thiooxidans[J].J Environ Sci,2006,26(12):2000-2007]
22 Solicio C,Lodi A,Veglio F.Bioleaching of zinc and aluminium from industrial waste sludge by means of Thiobacillus ferrooxidans[J].Waste Mana,2002,22:667-675