绿脓杆菌螯铁蛋白分泌及其对铜绿假单胞菌NY3降解烃类的影响作用
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摘要
为研究铜绿假单胞菌NY3胞外分泌物PCH对菌体自身及其对烃类降解的作用,对PCH影响烃类降解的效率及机理进行研究。分别以NY3菌降解烃类污染物的无机盐培养基和M9培养基为例,研究PCH含量对NY3菌降解十六烷活性的影响。结果表明,PCH分泌量与对NY3菌生长量及其对十六烷的比降解率呈负相关。将PCH加入到NY3菌降解十六烷体系中,发现少量PCH对NY3菌生长及其十六烷降解效率有促进作用,PCH浓度超过5.2 mg/L时,十六烷降解率及菌体的生长明显被抑制。进一步研究发现,PCH浓度过高会对NY3菌自身造成一定损害,抑制NY3菌烷氧化酶活性,同时增强降解体系中自由基信号强度,从而导致细胞对十六烷降解能力下降,同时使细胞大量死亡。该研究结果表明,使用NY3菌修复石油烃污染物过程中应严格控制PCH分泌量,为NY3菌在石油烃修复的应用提供参考。
In order to study the effect of pyochelin( PCH) of extracellular secretion of Pseudomonas aeruginosa NY3 on thalamus itself and hydrocarbons degradation,the efficiency and mechanism of the effects on hydrocarbons degradation of PCH were studies. Medium with inorganic salt of pollutant of hydrocarbons degraded by NY3 and M9 medium were taken as examples respectively to study the effects of PCH content on the activity of NY3 to degrade cetane. The results showed that PCH secretion assumed negative correlation with the NY3 growth and hexadecane specific degradation efficiency. PCH was added into cetane degradation system of NY3,it was found that a small amount of PCH could promote the growth of NY3 and hexadecane degradation efficiency. However,when the concentration of PCH was over 5. 2 mg/L,both the degradation and the growth rate were obviously suppressed. Further studies found that over-high PCH concentration may cause damage to NY3,inhibiting NY3 alkanes oxidase activity at the same time enhancing free radical signal intensity in the degradation system,and resulted in a decline of the cell degradation ability while causing cells to die in large number. The results of the study showed that the secretion of PCH should be rigidly controlled during the redemption process of petroleum hydrocarbon pollutant using NY3,and this provided an important theoretical guidance for the application of NY3 in the petroleum hydrocarbons' redemption.
In order to study the effect of pyochelin( PCH) of extracellular secretion of Pseudomonas aeruginosa NY3 on thalamus itself and hydrocarbons degradation,the efficiency and mechanism of the effects on hydrocarbons degradation of PCH were studies. Medium with inorganic salt of pollutant of hydrocarbons degraded by NY3 and M9 medium were taken as examples respectively to study the effects of PCH content on the activity of NY3 to degrade cetane. The results showed that PCH secretion assumed negative correlation with the NY3 growth and hexadecane specific degradation efficiency. PCH was added into cetane degradation system of NY3,it was found that a small amount of PCH could promote the growth of NY3 and hexadecane degradation efficiency. However,when the concentration of PCH was over 5. 2 mg/L,both the degradation and the growth rate were obviously suppressed. Further studies found that over-high PCH concentration may cause damage to NY3,inhibiting NY3 alkanes oxidase activity at the same time enhancing free radical signal intensity in the degradation system,and resulted in a decline of the cell degradation ability while causing cells to die in large number. The results of the study showed that the secretion of PCH should be rigidly controlled during the redemption process of petroleum hydrocarbon pollutant using NY3,and this provided an important theoretical guidance for the application of NY3 in the petroleum hydrocarbons' redemption.
引文
[1]Norman RS,Frontera-Suau R,Morris PJ.Variability in Pseudomonas aeruginosa lipopolysaccharide expression during crude oil degradation[J].Applied and Environmental Microbiology,2001,68(10):5096-5103.
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[3]马霞.铜绿假单胞菌NY3胞外活性物对其降解石油烃的作用研究[D].西安:西安建筑科技大学,2013.
[4]卢剑.鼠李糖脂在铜绿假单胞菌NY3代谢烃过程中的生理作用研究[D].西安:西安建筑科技大学,2014.
[5]徐鹏.石油烃代谢菌分泌绿脓菌素对其降解过程的影响作用研究[D].西安:西安建筑科技大学,2013.
[6]Sokol DA.Production and Utilization of Pyochelin by Clinical Isolated of Pseudomonas cepacia[J].Journal of Clinical Microbiology,1986,23(3):560-562.
[7]毛得奖,朱亚玲,韩宁.假单胞菌铁载体及色素研究[J].微生物学通报,2012,40(3):500-516.
[8]Britigan BE,Roeder TL,Rasmussen GT,et al.Interaction of the Pseudomonas aeruginosa Secretory Products Pyocyanin and Pyochelin Generates Hydroxyl Radical and Causes Synergistie Damage to Endothelial Cells[J].The Journal of Clinical Investigation,1992,90:2187-2196.
[9]Guo-Xin Sun,Jian-Jiang Zhong.Mechanism of Augmentation of Organotin Decomposition by Ferripyochelin:Formation of Hydroxyl Radical and Organotin-Pyochelin-Iron Ternary Complex[J].Applied and Environmental Microbiology,2006,72(11):7264-7269.
[10]De Witte JJ,Cox CD,Rasmussen GT,et al.Assessment of Structural Features of the Psedomonas Sideropore Pyochelin Required for Its Ability to Promote Oxidant-Mediated Endothelial Cell Injury[J].Archives Biochemistry and Biophsics,2001,393(2):236-244.
[11]孙红启.铁载体和铁离子对细菌生长过程的影响[D].济南:山东大学,2008.
[12]Nie M,Yin X,Ren C,et al.Novel rhamnolipid biosurfactants produced by a polycyclic aromatic hydrocarbon-degrading bacterium Pseudomonas aeruginosa strain NY3[J].Biotechnology Advances,2010,28(5):635-643.
[13]常虹,聂麦茜,葛碧洲,等.铜绿假单胞菌NY3所产表面活性剂对原油降解的影响[J].环境工程学报,2013,7(2):771-776.
[14]Heiko Hayen,Volmer DA.Volmer.Different iron-chelating properties of pyochelin diastereoisomers revealed by LC/MS[J].Anal Bioanal Chem,2006,385:606-611.
[15]Guo-Xin Sun,Wen-Qiang Zhou,Jian-Jiang Zhong.Organotin Decomposition by Pyochelin,Secreted by Psedomonas aeruginosa Even in an Iron-Sufficient Environment[J].Applied and Environmental Microbiology,2006,72(9):6411-6413.
[16]黄璐.铜绿假单胞菌NY3胞外分泌物对烃类降解的作用研究[D].西安:西安建筑科技大学,2015.
[17]林莹莹.固定化嗜油菌处理高含油废水及生物膜上自然配伍菌群结构特性研究[D].西安:西安建筑科技大学,2015.
[18]熊正英.自由基生物学在运动医学中的应用研究[J].陕西师范大学学报(自然科学版),2014,42(6):100-108.
[19]Cox CD,Grah am R.Isolation of an iron-binding compound from Pseudomonas aeruginosa[J].Journal of Bacteriology,1979,137(1):357-364.
[20]Youard ZA,Wenner N,Reimmann C.Iron acquisition with the natural siderophore enantiomers pyochelin and enantio-pyochelin in Pseudomonas species[J].Bio Metals,2011,24(3):513-522.
[2]常虹.铜绿假单胞菌NY3对高浓度油污泥液相处理技术研究[D].西安:西安建筑科技大学,2012.
[3]马霞.铜绿假单胞菌NY3胞外活性物对其降解石油烃的作用研究[D].西安:西安建筑科技大学,2013.
[4]卢剑.鼠李糖脂在铜绿假单胞菌NY3代谢烃过程中的生理作用研究[D].西安:西安建筑科技大学,2014.
[5]徐鹏.石油烃代谢菌分泌绿脓菌素对其降解过程的影响作用研究[D].西安:西安建筑科技大学,2013.
[6]Sokol DA.Production and Utilization of Pyochelin by Clinical Isolated of Pseudomonas cepacia[J].Journal of Clinical Microbiology,1986,23(3):560-562.
[7]毛得奖,朱亚玲,韩宁.假单胞菌铁载体及色素研究[J].微生物学通报,2012,40(3):500-516.
[8]Britigan BE,Roeder TL,Rasmussen GT,et al.Interaction of the Pseudomonas aeruginosa Secretory Products Pyocyanin and Pyochelin Generates Hydroxyl Radical and Causes Synergistie Damage to Endothelial Cells[J].The Journal of Clinical Investigation,1992,90:2187-2196.
[9]Guo-Xin Sun,Jian-Jiang Zhong.Mechanism of Augmentation of Organotin Decomposition by Ferripyochelin:Formation of Hydroxyl Radical and Organotin-Pyochelin-Iron Ternary Complex[J].Applied and Environmental Microbiology,2006,72(11):7264-7269.
[10]De Witte JJ,Cox CD,Rasmussen GT,et al.Assessment of Structural Features of the Psedomonas Sideropore Pyochelin Required for Its Ability to Promote Oxidant-Mediated Endothelial Cell Injury[J].Archives Biochemistry and Biophsics,2001,393(2):236-244.
[11]孙红启.铁载体和铁离子对细菌生长过程的影响[D].济南:山东大学,2008.
[12]Nie M,Yin X,Ren C,et al.Novel rhamnolipid biosurfactants produced by a polycyclic aromatic hydrocarbon-degrading bacterium Pseudomonas aeruginosa strain NY3[J].Biotechnology Advances,2010,28(5):635-643.
[13]常虹,聂麦茜,葛碧洲,等.铜绿假单胞菌NY3所产表面活性剂对原油降解的影响[J].环境工程学报,2013,7(2):771-776.
[14]Heiko Hayen,Volmer DA.Volmer.Different iron-chelating properties of pyochelin diastereoisomers revealed by LC/MS[J].Anal Bioanal Chem,2006,385:606-611.
[15]Guo-Xin Sun,Wen-Qiang Zhou,Jian-Jiang Zhong.Organotin Decomposition by Pyochelin,Secreted by Psedomonas aeruginosa Even in an Iron-Sufficient Environment[J].Applied and Environmental Microbiology,2006,72(9):6411-6413.
[16]黄璐.铜绿假单胞菌NY3胞外分泌物对烃类降解的作用研究[D].西安:西安建筑科技大学,2015.
[17]林莹莹.固定化嗜油菌处理高含油废水及生物膜上自然配伍菌群结构特性研究[D].西安:西安建筑科技大学,2015.
[18]熊正英.自由基生物学在运动医学中的应用研究[J].陕西师范大学学报(自然科学版),2014,42(6):100-108.
[19]Cox CD,Grah am R.Isolation of an iron-binding compound from Pseudomonas aeruginosa[J].Journal of Bacteriology,1979,137(1):357-364.
[20]Youard ZA,Wenner N,Reimmann C.Iron acquisition with the natural siderophore enantiomers pyochelin and enantio-pyochelin in Pseudomonas species[J].Bio Metals,2011,24(3):513-522.