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The catalytic degradation mechanism of petroleum hydrocarbons by Ferripyochelin (FerriPCH), formed by the combination of pyochelin (PCH) from the extracellular fluid of P. aeruginosa NY3 and Ferri, was investigated. The results showed that two chiral PCHs were produced by strain NY3. The purified PCH could degrade both alkanes and polycyclic aromatic compounds of petroleum hydrocarbon after combining with Ferri. A certain ratio of hexadecane, anthracene, as well as phenanthrene could be degraded by FerriPCH which formed by PCH and Ferri in the ratio from 2:1 to 1:1. In addition, degradation efficiency of aromatic hydrocarbons with low bio-availability was significantly higher than that of alkanes. Further studies showed that strongly oxidizing free radicals of •OH and •O2- produced after the binding of Ferri and PCH degraded petroleum hydrocarbons into carbon dioxide and water step by step.

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探讨了铜绿假单胞菌Pseudomonas aeruginosa NY3分泌的螯铁蛋白(Pyochelin,PCH)与铁形成复合物(Ferripyochelin,FerriPCH)催化降解石油烃的作用机理.结果表明,菌株NY3分泌的2种PCH为手性对映体.当纯化后的PCH与Fe3+鳌合时,可降解石油烃中的烷烃和多环芳烃等污染物.当PCH与Fe3+的比例为2:1 ~ 1:1时,对十六烷,蒽和菲有一定的降解效果,且生物利用度低的芳香烃降解效率显著高于烷烃.研究发现,Fe3+与PCH鳌合会产生•OH和•O2-自由基,这些具有强氧化性能的自由基可将石油烃逐步降解为二氧化碳和水.

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* 责任作者,讲师,
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黄芷若(2000-),女,湖北宜昌人,西安建筑科技大学硕士研究生,主要研究方向为难降解有机污染物的微生物处理技术..

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黄芷若(2000-),女,湖北宜昌人,西安建筑科技大学硕士研究生,主要研究方向为难降解有机污染物的微生物处理技术..

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黄芷若(2000-),女,湖北宜昌人,西安建筑科技大学硕士研究生,主要研究方向为难降解有机污染物的微生物处理技术..

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Chemical Engineering Journal2024502:157918., articleTitle=Comprehensive evaluation and Mechanistic comparison of Cr-Catalyzed homogeneous Fenton-Like reactions for coexisting organics degradation, refAbstract=null), Reference(id=1241408738905609105, tenantId=1146029695717560320, journalId=1234093305789726721, articleId=1241408718538068037, doi=null, pmid=null, pmcid=null, year=2024, volume=496, issue=null, pageStart=null, pageEnd=null, url=null, language=null, rfNumber=[27], rfOrder=26, authorNames=Cao Z Z, Xu J L, Dai J N, journalName=Chemical Engineering Journal, refType=null, unstructuredReference=Cao Z ZXu J LDai J N,et al. Direct oxidation of adsorbed petroleum hydrocarbons by activating persulfate with Fe/Mn-SOM complex performed in-situ for heavily contaminated soil remediation [J]. Chemical Engineering Journal2024496:154049., articleTitle=Direct oxidation of adsorbed petroleum hydrocarbons by activating persulfate with Fe/Mn-SOM complex performed in-situ for heavily contaminated soil remediation, refAbstract=null), Reference(id=1241408739006272408, tenantId=1146029695717560320, journalId=1234093305789726721, articleId=1241408718538068037, doi=null, pmid=null, pmcid=null, year=2021, volume=404, issue=null, pageStart=12568, pageEnd=null, url=null, language=null, rfNumber=[28], rfOrder=27, authorNames=Bokam R, Indumathi M N, Suresh K G, journalName=Chemical Engineering Jouranl, refType=null, unstructuredReference=Bokam RIndumathi M NSuresh K G. Investigating the degradation of nC12to nC23alkanes and PAHs in petroleum- contaminated water by electrochemical advanced oxidation process using an inexpensive Ti/Sb-SnO2/PbO2 anode [J]. Chemical Engineering Jouranl2021404:12568., articleTitle=Investigating the degradation of nC12to nC23alkanes and PAHs in petroleum- contaminated water by electrochemical advanced oxidation process using an inexpensive Ti/Sb-SnO2/PbO2 anode, refAbstract=null), Reference(id=1241408739136295841, tenantId=1146029695717560320, journalId=1234093305789726721, articleId=1241408718538068037, doi=null, pmid=null, pmcid=null, year=2023, volume=448, issue=null, pageStart=null, pageEnd=null, url=null, language=null, rfNumber=[29], rfOrder=28, authorNames=Zhang T, Chen Y, Wang T, journalName=Journal of Hazardous Materials, refType=null, unstructuredReference=Zhang TChen YWang T,et al. Efficient removal of petroleum hydrocarbons from soil by percarbonate with catechin-promoted Fe(III)/Fe(II) redox cycling: Activation of ferrous and roles of ⋅OH and⋅CO3 [J]. 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(a)PCH紫外-可见吸收光谱图;(b)PCH液相色谱图;(c)PCH正离子一级质谱图;(d)PCH正离子二级质谱图

, figureFileSmall=+YWfr4Vy5DeLFLYDMi2+Gg==, figureFileBig=7X6YvcZCTXEoE+9hakdmnw==, tableContent=null), ArticleFig(id=1241408732844839369, tenantId=1146029695717560320, journalId=1234093305789726721, articleId=1241408718538068037, language=EN, label=Fig.2, caption=Effect of FerriPCH on the degradation efficiency of hexadecane, figureFileSmall=/pqS2xOpE7cpskyo5LYjiQ==, figureFileBig=skZZa60P4aGeApc5fnMz5Q==, tableContent=null), ArticleFig(id=1241408733058748896, tenantId=1146029695717560320, journalId=1234093305789726721, articleId=1241408718538068037, language=CN, label=图2, caption=FerriPCH对十六烷降解效率的影响

(a)PCH质量浓度对十六烷降解效率的影响;(b)Fe3+质量浓度对十六烷降解效率的影响;(c)H2O2浓度对FerriPCH降解十六烷效率的影响

, figureFileSmall=/pqS2xOpE7cpskyo5LYjiQ==, figureFileBig=skZZa60P4aGeApc5fnMz5Q==, tableContent=null), ArticleFig(id=1241408733255881195, tenantId=1146029695717560320, journalId=1234093305789726721, articleId=1241408718538068037, language=EN, label=Fig.3, caption=Effect of FerriPCH on the degradation feeiciency of anthracene and phenanthrene, figureFileSmall=4x5Kdovtd5h0d/3uw0qj6Q==, figureFileBig=c6ObiJW+DRzRs3AZLQBjZA==, tableContent=null), ArticleFig(id=1241408733411070457, tenantId=1146029695717560320, journalId=1234093305789726721, articleId=1241408718538068037, language=CN, label=图3, caption=FerriPCH对蒽和菲降解效率的影响

(a)PCH质量浓度对蒽降解效率的影响;(b)PCH质量浓度对菲降解效率的影响

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Intermediates produced in the degradation of petroleum hydrocarbons by FerriPCH

, figureFileSmall=null, figureFileBig=null, tableContent=
序号石油烃主要离子(m/z)结构式名称
1十六烷146,131,103,89,75,73,61,59,45正丁醇
2174,159,143,128,114,100,86,73,56,454,5-二羟基-2-戊烯
3174,159,144,130,86,73,58,452-甲基-3羟基戊烷
4204,189,174,130,100,86,75,59,456-羟基己酸
5188,173,158,144,102,73,59,452-甲基-3-羟基己烷
6290,275,188,170,147,131,98,73己二酸
7328,313,201,132,117,73,55棕榈酸
8247,172,147,129,75,73,55,45琥珀酸
9234,203,189,147,144,129,103,731,2-二羟基萘
10306,216,176,147,128,102,731,2-二羟基-3,4-二氢萘
11331,275,227,172,147,129,111,731-羟基-2-(3-羟基-1-丙烯基)-3,4-二氢萘
12227,191,143,129,111,98,75,55,412-羧甲基-3-羟甲基-1,2-二氢萘
13246,216,147,103,73,552-羧基-1,2-二氢萘
14306,216,176,147,128,102,732,3-二羟基-1,2-二氢萘
15208,180,152,126,76,509,10-蒽醌
), ArticleFig(id=1241408734191211061, tenantId=1146029695717560320, journalId=1234093305789726721, articleId=1241408718538068037, language=CN, label=表1, caption=

FerriPCH降解石油烃过程中产生的中间产物

, figureFileSmall=null, figureFileBig=null, tableContent=
序号石油烃主要离子(m/z)结构式名称
1十六烷146,131,103,89,75,73,61,59,45正丁醇
2174,159,143,128,114,100,86,73,56,454,5-二羟基-2-戊烯
3174,159,144,130,86,73,58,452-甲基-3羟基戊烷
4204,189,174,130,100,86,75,59,456-羟基己酸
5188,173,158,144,102,73,59,452-甲基-3-羟基己烷
6290,275,188,170,147,131,98,73己二酸
7328,313,201,132,117,73,55棕榈酸
8247,172,147,129,75,73,55,45琥珀酸
9234,203,189,147,144,129,103,731,2-二羟基萘
10306,216,176,147,128,102,731,2-二羟基-3,4-二氢萘
11331,275,227,172,147,129,111,731-羟基-2-(3-羟基-1-丙烯基)-3,4-二氢萘
12227,191,143,129,111,98,75,55,412-羧甲基-3-羟甲基-1,2-二氢萘
13246,216,147,103,73,552-羧基-1,2-二氢萘
14306,216,176,147,128,102,732,3-二羟基-1,2-二氢萘
15208,180,152,126,76,509,10-蒽醌
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铜绿假单胞菌螯铁蛋白催化降解石油烃的机理
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黄芷若 1 , 郭镝妮 1 , 赵涛 1 , 聂红云 1, 2, * , 聂麦茜 1, 2 , 王磊 1, 2 , 赵碧洁 1
中国环境科学 | 环境微生物 2025,45(4): 2298-2304
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中国环境科学 | 环境微生物 2025, 45(4): 2298-2304
铜绿假单胞菌螯铁蛋白催化降解石油烃的机理
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黄芷若1 , 郭镝妮1, 赵涛1, 聂红云1, 2, * , 聂麦茜1, 2, 王磊1, 2, 赵碧洁1
作者信息
  • 1.西安建筑科技大学环境与市政工程学院,陕西 西安 710055
  • 2.陕西省膜分离重点实验室,陕西 西安 710055
  • 黄芷若(2000-),女,湖北宜昌人,西安建筑科技大学硕士研究生,主要研究方向为难降解有机污染物的微生物处理技术..

通讯作者:

* 责任作者,讲师,
The catalytic degradation mechanism of petroleum hydrocarbons by Ferripyochelin produced from the extracellular fluid of Pseudomonas aeruginosa NY3
Zhi-ruo HUANG1 , Di-ni GUO1, Tao ZHAO1, Hong-yun NIE1, 2, * , Mai-qian NIE1, 2, Lei WANG1, 2, Bi-jie ZHAO1
Affiliations
  • 1.School of Environmental and Municipal Engineering, Xi’an University of Architecture and Technology, Xi’an 710055, China
  • 2.Key Laboratory of Membrane Separation, Xi’an 710055, China
出版时间: 2025-04-20
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探讨了铜绿假单胞菌Pseudomonas aeruginosa NY3分泌的螯铁蛋白(Pyochelin,PCH)与铁形成复合物(Ferripyochelin,FerriPCH)催化降解石油烃的作用机理.结果表明,菌株NY3分泌的2种PCH为手性对映体.当纯化后的PCH与Fe3+鳌合时,可降解石油烃中的烷烃和多环芳烃等污染物.当PCH与Fe3+的比例为2:1 ~ 1:1时,对十六烷,蒽和菲有一定的降解效果,且生物利用度低的芳香烃降解效率显著高于烷烃.研究发现,Fe3+与PCH鳌合会产生•OH和•O2-自由基,这些具有强氧化性能的自由基可将石油烃逐步降解为二氧化碳和水.

螯铁蛋白  /  FerriPCH  /  催化降解  /  石油烃  /  自由基

The catalytic degradation mechanism of petroleum hydrocarbons by Ferripyochelin (FerriPCH), formed by the combination of pyochelin (PCH) from the extracellular fluid of P. aeruginosa NY3 and Ferri, was investigated. The results showed that two chiral PCHs were produced by strain NY3. The purified PCH could degrade both alkanes and polycyclic aromatic compounds of petroleum hydrocarbon after combining with Ferri. A certain ratio of hexadecane, anthracene, as well as phenanthrene could be degraded by FerriPCH which formed by PCH and Ferri in the ratio from 2:1 to 1:1. In addition, degradation efficiency of aromatic hydrocarbons with low bio-availability was significantly higher than that of alkanes. Further studies showed that strongly oxidizing free radicals of •OH and •O2- produced after the binding of Ferri and PCH degraded petroleum hydrocarbons into carbon dioxide and water step by step.

pyochelin  /  FerriPCH  /  catalytic degradation  /  petroleum hydrocarbons  /  free radicals
黄芷若, 郭镝妮, 赵涛, 聂红云, 聂麦茜, 王磊, 赵碧洁. 铜绿假单胞菌螯铁蛋白催化降解石油烃的机理. 中国环境科学, 2025 , 45 (4) : 2298 -2304 .
Zhi-ruo HUANG, Di-ni GUO, Tao ZHAO, Hong-yun NIE, Mai-qian NIE, Lei WANG, Bi-jie ZHAO. The catalytic degradation mechanism of petroleum hydrocarbons by Ferripyochelin produced from the extracellular fluid of Pseudomonas aeruginosa NY3[J]. China Environmental Science, 2025 , 45 (4) : 2298 -2304 .
在石油开采,储存,运输和加工过程中产生的石油烃,已成为土壤,水和大气环境的主要有机污染物之一[1].由于其成分对低级和高级生物均具有的致癌性,致畸性,诱变性和免疫毒性,石油污染处理技术已成为全球关注的焦点问题[2].从环境可持续发展来看,生物修复相较于物理和化学处理技术更前景[3].尽管生物法具有环保,低成本的优势,但其受环境因素,石油污染物的毒性以及较长修复周期等因素的制约[4-5].由于环境因素可通过优化特定处理工艺来满足,因此提高微生物对石油烃,特别是难降解组分,如多环芳烃的降解效率已成为研究热点[6,8].
近几十年里,生物体系中的自由基降解在处理难降解有机污染物(ROPs)中逐渐被更多学者认可[9,12].在自由基降解途径中,通过胞外活性化合物催化产生H2O2,O2-和•OH[13]等活性氧(ROS),这些具有氧化还原活性的ROS进一步将母体污染物转化为低毒性的,可生物降解的中间产物[9],从而提高ROPs的总生物降解效率.因此,揭示自由基降解的机制对提高ROPs的生物利用度至关重要.目前,已报道了多种能形成ROS的胞外活性化合物.其中,由铜绿假单胞菌和伯克氏菌分泌的螯铁蛋白(Pyochelin,PCH)[14-15]是胞外活性化合物之一.当PCH与铁离子结合时,会在细胞外液中形成具有ROS生成能力的FerriPCH.已有研究探讨了FerriPCH对四溴双酚A等阻燃剂的降解作用[11,13].铜绿假单胞菌和伯克氏菌作为分泌PCH的主要菌株,在石油烃污染场地的修复中得到广泛的应用[3,16],但FerriPCH对烷烃,多环芳烃等石油烃的降解机制尚不清楚.
本文研究了FerriPCH对石油烃中十六烷和多环芳烃(蒽和菲)的降解机制.PCH是由铜绿假单胞菌分泌的铁载体,通过从细胞外液中提取并采用UV-Vis光谱法和LC-MS进行表征;并测定FerriPCH对十六烷,蒽和菲的降解效率.同时,通过鉴定代谢产物以及自由基降解过程中产生的自由基以揭示其降解机理.
铜绿假单胞菌NY3分离于陕西石油污染土壤[17],并对其做16rRNA鉴定,结果提交NCBI数据库,登录号为GU377209.
牛肉膏培养基:根据参考文献[18]制备.组成为NaCl 5g/L,蛋白胨10g/L,牛肉膏3g/L,调pH值至7.8,121℃灭菌30min,备用.
M9培养基:采用M9培养基制备PCH[19].组成为Na2HPO4 64.0g,KH2PO4 15.0g,NaCl 2.5g,NH4Cl 5.0g,丁二酸钠20mmol/L,调pH值至7.8,121℃灭菌30min,备用.
将单菌落NY3菌株接种于牛肉膏培养基中,于31℃,150r/min恒温摇床中震荡培养.培养好的种子液以10%的接种量(OD600nm=1.60±0.08)接种到M9培养基中,并添加20mmol/L琥珀酸钠作为唯一碳源,于31℃,150r/min好氧培养3d,10000r/min离心15min去除菌体,得到无菌胞外液,用6mol/L的盐酸调pH值至2.5.采用文献[19]报道的方法提取上清液中的PCH.100mL乙酸乙酯液萃取上清液3次后,用旋转蒸发仪去除溶剂.乙酸乙酯挥发后的残留物溶于1mL甲醇/水(1:2,体积比)中,保留在固相萃取柱中(SupelcleanTM ENVITM-18).分别用5mL甲醇/水(1:20,体积比)和5mL甲醇/水(1:1,体积比)经行清洗,然后用10mL 2%甲酸洗脱.洗脱液在氮气气流下浓缩,即得到纯化的PCH.
用紫外-可见分光光度计测量PCH的吸收光谱.将纯化的PCH溶解在超纯水中制备样品,以超纯水作为空白对照.扫描范围设置为200~800nm.
用高效液相-离子阱-飞行时间质谱(Shimadzu Corporation. Kyoto,Japan)检测提取到的PCH[20].流动相体系由含有0.1%甲酸(体积分数)的超纯水(溶剂A)和高效液相色谱级乙腈(溶剂B)组成.洗脱程序以20% B开始,稳定1min,在20min内从20% B到95% B进行线性梯度洗脱.溶剂B的浓度上升至95%后,保持5min,随后返回到20% B的初始流动相条件.其中最佳进样量为20μL,流速设定为0.2mL/min.PCH在C18色谱柱(Shimpack XR-ODS,2.2μm总孔,2.0mm i.d×75mm;日本岛津公司)上实现分离.
选十六烷,蒽和菲为石油烃的典型代表.分别制备20mL的降解体系,并加入PCH,FeCl3,石油烃和Tris-HCl缓冲液(50mmol/L,pH=8.0).PCH和FeCl3浓度变化范围为0~400μmol/L,十六烷,蒽和菲的浓度分别为1.16g/L,25mg/L和25mg/L.当探究PCH浓度对FerriPCH降解体系影响时,Fe3+浓度设定为100μmol/L;当考察Fe3+浓度对FerriPCH降解体系影响时,PCH浓度为100μmol/L;探究H2O2对FerriPCH降解石油烃的影响时,H2O2,Fe3+的浓度均为100μmol/L.所有降解体系在31℃,151r/min条件下好氧培养.
十六烷的测定[10]:用5mL正己烷萃取降解体系中十六烷3次后,在配备有HP-5毛细管柱(5%phenyl Methyl Si-loxane,30m×320mm×0.25mm)和火焰电离检测器(FID)气相色谱分析仪上(安捷伦6890N,CA)进行定量分析.通过十六烷的初始浓度和终浓度计算十六烷的降解效率.
蒽和菲的提取和测定根据参考文献稍作修改[21]:降解体系中的蒽和菲用5mL环己烷萃取3次后,合并有机相并蒸发有机溶剂.将固体溶解在5mL色谱级甲醇中,用液相色谱(Thermo,U3000)对降解体系中残留的蒽和菲进行定量分析.进样量为20μL,流速为1.0mL/min,检测器波长为254nm.通过十六烷的初始浓度和最终浓度计算蒽和菲的降解效率.
中间产物的提取和鉴定如下[22]:降解体系在10000r/min条件下离心10min后收集上清液.对于酸性中间产物,上清液用6mol/L HCl调节pH值至2~3,并用10mL乙酸乙酯萃取3次.对于碱性中间产物,用10mmol/L NaOH溶液对酸性馏分中的乙酸乙酯萃取3次.随后,用6mol/L HCl将氢氧化钠相酸化至pH值2~3,用10mL乙酸乙酯萃取3次.最后,将乙酸乙酯相干燥,浓缩后,用DB-5色谱柱(安捷伦,7000B)进行气相色谱-质谱分析.程序设置为:氦气为流动相,流速1.0mL/min,进样口温度为280℃,进样量为1μL,源离子温度为230℃,升温程序:160℃保持1min,然后以20℃/min升温至260℃,保持6min.
待检测溶液体系总体积为50μL,其中DMPO,PCH和FeCl3的浓度分别为200,100和100μmol/L,最后用Tris-HCl缓冲液(50mmol/L,pH=8.0)调整至所需体积.EPR测量使用Bruker 169EMXmicro光谱仪(Bruker BioSpin,Billerica,MA,USA)[10].EMXmicro光谱仪参数如下:微波功率20mW,调制频率100kHz,调制幅度1.0G,时间常数0.327或0.655s,放大倍数5.0×105.
除气相色谱-质谱联用仪测量外,本工作中所有实验均至少重复3次,用Origin 2022软件对实验数据进行标准差分析.
H2O2对FerriPCH降解十六烷,蒽和菲的显著性分析:利用SPSS软件(IBM SPSS Statistics 27)对加H2O2体系和对照组做独立样本T检验,分析两者的显著性,P<0.05说明具有显著性.(用∗表示相关性强弱,∗∗∗:P <0.001; ∗∗:P<0.01; ∗:0.01<P<0.05).
PCH是铜绿假单胞菌分泌的一种铁载体.在M9培养基中发酵培养后,NY3菌株可分泌64~70mg/L的PCH.采用1.3的方法对其提取纯化,可获得50%以上的PCH粗品,其色谱纯度约为49.3%.采用紫外-可见光谱法,液相色谱-质谱法对PCH进行表征,结果如图1所示.从图1a可以看出,纯化得到的PCH在216,253,288,327和360nm处分别有5个吸收峰.其中,在216,253和327nm处的峰与P.cepaciaP.aeruginosa PAO1[23]的螯铁蛋白紫外吸收峰相同.LC-MS(图1b-d)结果显示,液相色谱谱图(图1b)中出现的两个吸收峰,其分子量均为324(图1c),二级质谱(图1d)中出现的m/z 281,m/z 247,m/z 224,m/z 190,m/z 172和m/z 186的碎片离子应为[M+H-COO],[M+H-OH-S-CH=CH-CH=CH],[M+H-COO H-S-N-CH3-CH-H],[M+H-COOH-N-S-S-OH-CH]和[M+H-C6H4-OH-C-S-N-CH2-CH-H]的离子碎片.这与铜绿假单胞菌ATCC15692[24]产生的PCH质谱图相同.该结果表明NY3菌株产生的PCH由两个分子量相同的手性对映体组成.
图2a可知,当PCH浓度为0,Fe3+浓度为100μmol/L时,十六烷几乎不降解;随着PCH浓度增加,十六烷降解率逐步增加,当PCH浓度为50和100时,十六烷降解率最高.由图2b可知,当Fe3+浓度为0,PCH浓度为100μmol/L时,十六烷几乎不降解;随着体系中Fe3+浓度增加,十六烷降解率显著增加.以上结果表明Fe3+和PCH单独存在均不能降解十六烷,但当Fe3+与PCH螯合形成FerriPCH后可引起十六烷降解,且当PCH与Fe3+的浓度比为2:1至1:1之间时,十六烷的降解效率较高,这与之前所报道PCH:Fe3+复合物的最优化学计量配比一致[24].但总体的降解率不高,约为13.8%左右.由前人研究结果可知,虽然H2O2不能引起有机污染物的降解[25-26],但与Fe3+结合可迅速降解有机污染物.因此在FerriPCH体系中加入H2O2,其对十六烷降解率的影响如图2c所示.与FerriPCH体系相比,加入过氧化氢对十六烷降解率无显著促进;但H2O2加入后,十六烷的降解效率在更宽的PCH浓度范围内保持在相对较高的水平.综上可知,PCH与Fe3+螯合能引起十六烷降解,但降解率低;过氧化氢的加入并未提高十六烷降解率.
保持实验条件与十六烷降解体系一致,考察了FerriPCH对蒽和菲等芳烃的降解.由图3可知,在FerriPCH体系中加入H2O2时,蒽和菲的降解效率得到提高,随着PCH浓度的增加,当Fe3+与PCH的比例在2:1至1:1范围内时,FerriPCH对蒽和菲的降解效果最佳.最优条件下,蒽的降解率可达25%左右,菲的降解率可达50%左右.相较于十六烷降解体系,降解效率分别提升11.76%和41.14%.这与Cao等[27]在Fe/Mn-SOM复合物体系中降解烷烃和芳香烃的结论一致.究其原因,可能源于烷烃和芳香烃分子结构的根本差异.芳香烃中交替的C=C双键增强了与活性氧自由基的反应活性,且C=C键断裂所需能量比长链烷烃中C-C键低[28],从而致使生物利用度低的芳香烃反而更易被FerriPCH催化降解.此现象利于微生物对芳香烃的高效降解,进而提高石油烃的总降解率.
采用气相色谱-质谱法确定了FerriPCH催化下自由基降解过程中产生的中间产物.中间产物通过将其质谱特征峰与NIST库数据库的标准谱图或与已发表的质谱图比较进行鉴定,结果列于表1.如表1所示,十六烷降解体系的中间产物包括棕榈酸,己二酸,2-甲基-3-羟基己烷,6-羟基己酸,2-甲基-3-羟基戊烷,4,5-二羟基-2-戊烯和正丁醇.菲降解的中间产物包括1-羟基-2-(3-羟基-1-丙烯)-1,2,3,4-四氢萘,1,2-羟基-3,4-二氢萘,1,2-二羟基萘和琥珀酸.蒽降解的中间产物包括9,10-蒽醌,3,4-二羟基-2,3-二氢萘,2-羧甲基-1,2-二羟基萘,2-羧甲基-3-羟乙基-1,2-二氢.烷烃和芳香烃的降解途径涉及活性氧的介导自由基反应[29]:经氧化,开环或裂解,生成包括醇类,羧酸等中间产物,这些中间产物最终被彻底氧化成CO2和H2O.上述结果表明,在FerriPCH催化作用下,十六烷,蒽和菲等逐步降解为碳原子数减少的中间产物,进而被矿化.不同石油烃的降解途径如图4所示.
用电子顺磁共振(EPR)检测石油烃降解过程中产生的活性自由基.EPR谱如图5所示.在Tris-HCl缓冲液中仅加入DMPO,PCH和DMPO,或加入Fe3+和DMPO,均未检测到明显信号.说明PCH和Fe3+单独存在的情况下都不会产生自由基.当Fe3+加入到PCH和DMPO的体系中时,观察到特征性的DMPO−•OH (aN=aH=14.9G)信号,证实了Fe3+与PCH螯合过程中•OH的产生.为进一步验证Fe3+在反应中的作用,向体系中加入100mmol/L乙二胺四乙酸二钠作为Fe3+的捕获剂,结果无信号产生,表明Fe3+是促进•OH自由基生成的必要条件.异丙醇和苯醌是•OH和超氧阴离子自由基(•O2-)高效捕获剂.当在Fe3+,PCH和DMPO的Tris-HCl体系中分别加入异丙醇和苯醌时,信号均未完全消失,表明Fe3+和PCH的螯合过程中不仅产生了•OH,而且产生了•O2-.在其体系中加入H2O2,能极大的促进体系中•OH自由基的浓度.据了解,这是首次在Fe3+与PCH体系中检测到•O2-存在[30].根据石油烃的降解效率可知,加入过氧化氢有效提升FerriPCH体系中强氧化性•OH的浓度.基于密度泛函理论,多环芳烃因其亲电反应指数高,更易受到•OH等自由基的攻击[27],因此,蒽和菲的降解效率高于十六烷.
3.1 铜绿假单胞菌NY3菌株分泌的PCH主要由两种手性对映体组成.当PCH与Fe3+以物质的量浓度比为2:1~1:1形成FerriPCH复合物时,对石油烃的降解效率最高.
3.2 相较于十六烷,FerriPCH复合物对难以生物降解的蒽和菲的降解效率更高.在FerriPCH的催化降解作用下,十六烷,蒽和菲等石油烃类物质逐步降解,形成一系列碳原子数减少的中间产物,最终转化为二氧化碳和水.
3.3 降解机理的研究表明,当PCH和Fe3+鳌合形成FerriPCH复合物时,生成•OH和•O2-等自由基.加入H2O2,可显著促进体系中•OH的浓度.
  • 陕西省教育厅重点实验室项目(23JS040)
  • 陕西省秦岭生态安全重点实验室开放基金(QLES202501)
  • 陕西省科技创新团队计划(2024RS-CXTD-51)
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  • 接收时间:2024-09-11
  • 首发时间:2026-03-19
  • 出版时间:2025-04-20
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  • 收稿日期:2024-09-11
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陕西省教育厅重点实验室项目(23JS040)
陕西省秦岭生态安全重点实验室开放基金(QLES202501)
陕西省科技创新团队计划(2024RS-CXTD-51)
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    1.西安建筑科技大学环境与市政工程学院,陕西 西安 710055
    2.陕西省膜分离重点实验室,陕西 西安 710055

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2种不同金属材料的力学参数

Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)

Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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