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Article(id=1208051025609597494, tenantId=1146029695717560320, journalId=1146123166801305609, issueId=1208051024368083510, articleNumber=null, orderNo=null, doi=10.12404/j.issn.1671-1815.2405989, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1723132800000, receivedDateStr=2024-08-09, revisedDate=1743436800000, revisedDateStr=2025-04-01, acceptedDate=null, acceptedDateStr=null, onlineDate=1765951409008, onlineDateStr=2025-12-17, pubDate=1751040000000, pubDateStr=2025-06-28, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1765951409008, onlineIssueDateStr=2025-12-17, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1765951409007, creator=13701087609, updateTime=1765951409007, updator=13701087609, issue=Issue{id=1208051024368083510, tenantId=1146029695717560320, journalId=1146123166801305609, year='2025', volume='25', issue='18', pageStart='7455', pageEnd='7883', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1765951408712, creator=13701087609, updateTime=1765951896766, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1208053071507198943, tenantId=1146029695717560320, journalId=1146123166801305609, issueId=1208051024368083510, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1208053071507198944, tenantId=1146029695717560320, journalId=1146123166801305609, issueId=1208051024368083510, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=7866, endPage=7873, ext={EN=ArticleExt(id=1208051026075165242, articleId=1208051025609597494, tenantId=1146029695717560320, journalId=1146123166801305609, language=EN, title=Adsorption Characteristics of Typical Heavy Metals by Sulfhydryl Groups in Natural Aquatic Biofilms, columnId=1156262729993277777, journalTitle=Science Technology and Engineering, columnName=Papers·Environmental and Safe Science, runingTitle=null, highlight=null, articleAbstract=

To determine the presence of sulfhydryl groups on natural aquatic biofilms and their adsorption characteristics for typical heavy metals, a method for sulfhydryl masking in the biofilms based on a specific masking agent was established in this study. Based on this method, surface concentration of sulfhydryl group in natural biofilms and their adsorption characteristics for typical heavy metals, including Cu, Pb, and Cd, at different pH were investigated. The results indicate that the established masking method can effectively mask sulfhydryl groups and has little effect on microorganisms in biofilms. There are relatively low concentrations of sulfhydryl groups on the surface of natural biofilms, with a concentration of (5.8 ± 0.6) μmol/g, accounting for 5.7% of the total site concentration on the biofilms. Despite the low concentration of sulfhydryl groups, their stronger metal binding capacity makes them significantly contribute to metal adsorption when the metal concentration is low (the theoretical loading of heavy metals by the biofilm is less than 1.0 μmol/g). This pattern is essentially unaffected by the pH of the adsorption system and the type of heavy metal. This proves that sulfhydryl groups in biofilms also have an important impact on the behavior and risk of heavy metals in natural aquatic environments with low metal content, further highlighting the environmental significance of natural biofilms.

, correspAuthors=Xiu-yi HUA, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, authorCompany=null, fund=null, authors=null, authorsList=Zhi-song ZHANG, Di-fei XIE, Yu GU, Xiu-yi HUA), CN=ArticleExt(id=1208051029489328789, articleId=1208051025609597494, tenantId=1146029695717560320, journalId=1146123166801305609, language=CN, title=自然水体生物膜巯基对典型重金属的吸附特征, columnId=1156262730140078420, journalTitle=科学技术与工程, columnName=论文·环境科学、安全科学, runingTitle=null, highlight=null, articleAbstract=

为了解地表水环境中广泛存在的自然水体生物膜表面巯基基团的存在情况和它们对典型重金属的吸附特征,建立了基于特异性巯基掩蔽剂的适用于自然水体生物膜中巯基掩蔽的方法,并利用该方法研究了生物膜中巯基的表面浓度,以及不同pH下生物膜巯基对典型重金属Cu、Pb和Cd的吸附特征。研究结果表明:所建立的巯基掩蔽方法能有效掩蔽巯基且对生物膜中微生物的影响很小。自然水体生物膜表面存在相对较低浓度的巯基基团,浓度为(5.8 ± 0.6) μmol/g,占生物膜表面总位点浓度的5.7%。尽管巯基浓度较低,但由于其具有更强的重金属结合能力,使其在重金属浓度较低时(吸附体系中生物膜对重金属的理论负载量 < 1.0 μmol/g时)对生物膜吸附重金属有重要贡献,显著影响生物膜吸附重金属的特征;且这种规律基本不受体系pH和重金属种类的影响。研究结果证明生物膜中巯基对天然水环境中重金属的环境行为和效应也有重要影响,进一步体现了自然水体生物膜的重要环境意义。

, correspAuthors=花修艺, authorNote=null, correspAuthorsNote=
* 花修艺(1976—),男,汉族,江苏邳州人,博士,教授。研究方向:环境化学。E-mail:。
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张之松(1969—),男,汉族,河北廊坊人,工程师。研究方向:环境管理和环境化学。E-mail:。

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张之松(1969—),男,汉族,河北廊坊人,工程师。研究方向:环境管理和环境化学。E-mail:。

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张之松(1969—),男,汉族,河北廊坊人,工程师。研究方向:环境管理和环境化学。E-mail:。

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figureFileSmall=ev9jmaZswPnmfV2RxLNJow==, figureFileBig=eZvowbWAHcvfAxN/UHoCIA==, tableContent=null), ArticleFig(id=1208085595742643083, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1208051025609597494, language=CN, label=图2, caption=不同浓度qBBr对生物膜中叶绿素a含量、SOD酶活性和MDA含量的影响, figureFileSmall=ev9jmaZswPnmfV2RxLNJow==, figureFileBig=eZvowbWAHcvfAxN/UHoCIA==, tableContent=null), ArticleFig(id=1208085595931386778, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1208051025609597494, language=EN, label=Fig.3, caption=The binding of qBBr to biofilms at different concentrations, figureFileSmall=Au58Uz4KI1wl3xQcunnz7Q==, figureFileBig=SgB8O+AVwkumAz7q6TVNiQ==, tableContent=null), ArticleFig(id=1208085596065604511, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1208051025609597494, language=CN, label=图3, caption=不同浓度qBBr与生物膜的结合量, figureFileSmall=Au58Uz4KI1wl3xQcunnz7Q==, figureFileBig=SgB8O+AVwkumAz7q6TVNiQ==, tableContent=null), 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ArticleFig(id=1208085597034488798, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1208051025609597494, language=EN, label=Fig.7, caption=The relationship between the difference of metal adsorption ratio of the biofilms before and after qBBr blocking at different pH and the theoretical metal loading, figureFileSmall=dNaOBjm6FHwjhz6uYn1BBg==, figureFileBig=SavQ3qlEO/7honnCIS+jUA==, tableContent=null), ArticleFig(id=1208085597181289451, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1208051025609597494, language=CN, label=图7, caption=不同pH下掩蔽前后生物膜对重金属吸附比例的差值与理论负载量的关系, figureFileSmall=dNaOBjm6FHwjhz6uYn1BBg==, figureFileBig=SavQ3qlEO/7honnCIS+jUA==, tableContent=null), ArticleFig(id=1208085597277758456, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1208051025609597494, language=EN, label=Table 1, caption=

pKa and concentration of the four apparent binding sites in the biofilms before and after qBBr blocking

, figureFileSmall=null, figureFileBig=null, tableContent=
表观结合
位点		 未掩蔽生物膜 qBBr掩蔽生物膜
pKa 浓度/
(μmol·g-1)		 pKa 浓度/
(μmol·g-1)
1 0.31±0.1 43.9±1.9 0.30±0.1 43.1±1.7
2 5.95±0.2 14.7±0.8 5.91±0.1 15.1±0.9
3 8.62±0.1 19.7±0.5 7.89±0.2 13.9±0.3
4 10.08±0.3 23.3±1.4 10.03±0.2 23.7±1.1
合计 — 101.6±1.6 — 95.8±1.2
), ArticleFig(id=1208085597386810368, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1208051025609597494, language=CN, label=表1, caption=

拟合得到的qBBr掩蔽前后生物膜的各表观结合点位的pKa及浓度

, figureFileSmall=null, figureFileBig=null, tableContent=
表观结合
位点		 未掩蔽生物膜 qBBr掩蔽生物膜
pKa 浓度/
(μmol·g-1)		 pKa 浓度/
(μmol·g-1)
1 0.31±0.1 43.9±1.9 0.30±0.1 43.1±1.7
2 5.95±0.2 14.7±0.8 5.91±0.1 15.1±0.9
3 8.62±0.1 19.7±0.5 7.89±0.2 13.9±0.3
4 10.08±0.3 23.3±1.4 10.03±0.2 23.7±1.1
合计 — 101.6±1.6 — 95.8±1.2
), ArticleFig(id=1208085597529415684, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1208051025609597494, language=EN, label=Table 2, caption=

The fitted parameters of Langmuir isotherm of Cu adsorption to the biofilms before and after qBBr blocking

, figureFileSmall=null, figureFileBig=null, tableContent=
pH 巯基 Γmax /(μmol·g-1) K/(L·μmol-1) R2
估计值 误差 估计值 误差
4.5 未掩蔽 8.96 1.78 0.222 0.056 0.998 3
掩蔽 12.51 5.77 0.129 0.070 0.996 5
5.2 未掩蔽 12.25 1.12 0.244 0.028 0.999 7
掩蔽 11.91 0.71 0.225 0.017 0.999 9
6.0 未掩蔽 13.31 1.61 0.502 0.084 0.998 7
掩蔽 13.47 0.53 0.433 0.024 0.999 9
7.0 未掩蔽 12.21 0.89 0.906 0.102 0.999 0
掩蔽 12.39 0.40 0.737 0.036 0.999 8
), ArticleFig(id=1208085597625884684, tenantId=1146029695717560320, journalId=1146123166801305609, articleId=1208051025609597494, language=CN, label=表2, caption=

qBBr掩蔽前后生物膜吸附Cu的Langmuir吸附等温线参数

, figureFileSmall=null, figureFileBig=null, tableContent=
pH 巯基 Γmax /(μmol·g-1) K/(L·μmol-1) R2
估计值 误差 估计值 误差
4.5 未掩蔽 8.96 1.78 0.222 0.056 0.998 3
掩蔽 12.51 5.77 0.129 0.070 0.996 5
5.2 未掩蔽 12.25 1.12 0.244 0.028 0.999 7
掩蔽 11.91 0.71 0.225 0.017 0.999 9
6.0 未掩蔽 13.31 1.61 0.502 0.084 0.998 7
掩蔽 13.47 0.53 0.433 0.024 0.999 9
7.0 未掩蔽 12.21 0.89 0.906 0.102 0.999 0
掩蔽 12.39 0.40 0.737 0.036 0.999 8
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自然水体生物膜巯基对典型重金属的吸附特征
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张之松 1, 2 , 谢涤菲 1, 3 , 谷裕 1 , 花修艺 1, *
科学技术与工程 | 论文·环境科学、安全科学 2025,25(18): 7866-7873
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科学技术与工程 | 论文·环境科学、安全科学 2025, 25(18): 7866-7873
自然水体生物膜巯基对典型重金属的吸附特征
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张之松1, 2 , 谢涤菲1, 3, 谷裕1, 花修艺1, *
作者信息
  • 1 吉林大学新能源与环境学院, 长春 130012
  • 2 国家管网集团建设项目管理分公司, 廊坊 065001
  • 3 吉林省长春生态环境监测中心, 长春 130012

    • 张之松(1969—),男,汉族,河北廊坊人,工程师。研究方向:环境管理和环境化学。E-mail:。

通讯作者:

* 花修艺(1976—),男,汉族,江苏邳州人,博士,教授。研究方向:环境化学。E-mail:。
Adsorption Characteristics of Typical Heavy Metals by Sulfhydryl Groups in Natural Aquatic Biofilms
Zhi-song ZHANG1, 2 , Di-fei XIE1, 3, Yu GU1, Xiu-yi HUA1, *
Affiliations
  • 1 College of New Energy and Environment, Jilin University, Changchun 130012, China
  • 2 Construction Project Management Branch, National Pipeline Network Group, Langfang 065001, China
  • 3 Changchun Ecological and Environmental Monitoring Centre of Jilin Province, Changchun 130012, China
出版时间: 2025-06-28 doi: 10.12404/j.issn.1671-1815.2405989
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摘要
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为了解地表水环境中广泛存在的自然水体生物膜表面巯基基团的存在情况和它们对典型重金属的吸附特征,建立了基于特异性巯基掩蔽剂的适用于自然水体生物膜中巯基掩蔽的方法,并利用该方法研究了生物膜中巯基的表面浓度,以及不同pH下生物膜巯基对典型重金属Cu、Pb和Cd的吸附特征。研究结果表明:所建立的巯基掩蔽方法能有效掩蔽巯基且对生物膜中微生物的影响很小。自然水体生物膜表面存在相对较低浓度的巯基基团,浓度为(5.8 ± 0.6) μmol/g,占生物膜表面总位点浓度的5.7%。尽管巯基浓度较低,但由于其具有更强的重金属结合能力,使其在重金属浓度较低时(吸附体系中生物膜对重金属的理论负载量 < 1.0 μmol/g时)对生物膜吸附重金属有重要贡献,显著影响生物膜吸附重金属的特征;且这种规律基本不受体系pH和重金属种类的影响。研究结果证明生物膜中巯基对天然水环境中重金属的环境行为和效应也有重要影响,进一步体现了自然水体生物膜的重要环境意义。

自然水体生物膜  /  巯基  /  吸附  /  铅  /  铜  /  镉

To determine the presence of sulfhydryl groups on natural aquatic biofilms and their adsorption characteristics for typical heavy metals, a method for sulfhydryl masking in the biofilms based on a specific masking agent was established in this study. Based on this method, surface concentration of sulfhydryl group in natural biofilms and their adsorption characteristics for typical heavy metals, including Cu, Pb, and Cd, at different pH were investigated. The results indicate that the established masking method can effectively mask sulfhydryl groups and has little effect on microorganisms in biofilms. There are relatively low concentrations of sulfhydryl groups on the surface of natural biofilms, with a concentration of (5.8 ± 0.6) μmol/g, accounting for 5.7% of the total site concentration on the biofilms. Despite the low concentration of sulfhydryl groups, their stronger metal binding capacity makes them significantly contribute to metal adsorption when the metal concentration is low (the theoretical loading of heavy metals by the biofilm is less than 1.0 μmol/g). This pattern is essentially unaffected by the pH of the adsorption system and the type of heavy metal. This proves that sulfhydryl groups in biofilms also have an important impact on the behavior and risk of heavy metals in natural aquatic environments with low metal content, further highlighting the environmental significance of natural biofilms.

natural biofilm  /  sulfhydryl group  /  adsorption  /  lead  /  copper  /  cadmium
张之松, 谢涤菲, 谷裕, 花修艺. 自然水体生物膜巯基对典型重金属的吸附特征. 科学技术与工程, 2025 , 25 (18) : 7866 -7873 . DOI: 10.12404/j.issn.1671-1815.2405989
Zhi-song ZHANG, Di-fei XIE, Yu GU, Xiu-yi HUA. Adsorption Characteristics of Typical Heavy Metals by Sulfhydryl Groups in Natural Aquatic Biofilms[J]. Science Technology and Engineering, 2025 , 25 (18) : 7866 -7873 . DOI: 10.12404/j.issn.1671-1815.2405989
正文
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重金属是水环境中一类具有显著环境风险的重要污染物,受到了广泛关注。各种固相物质对重金属的吸附是影响重金属水环境行为的最重要因素之一[1-2]。水环境中广泛存在的、由水中各种微生物和有机-无机非生物成分组成的天然生物膜是一类会显著影响重金属等污染物行为的重要活性物质[3-5]。作为典型的复杂固相物质,生物膜中的不同化学成分和基团对重金属的吸附有不同的贡献和复杂的影响:微观上,生物膜中微生物、有机物和无机成分表面广泛存在的羟基、羧基、氨基、磷酰基、巯基等基团会通过表面配合等作用结合重金属,从而影响生物膜对重金属的宏观吸附作用,改变重金属的行为[2,6-8]。
硫(-II)与过渡金属之间往往有很强的结合作用,如金属硫化物往往有极低的溶度积、巯基(-SH)往往可以与重金属强烈结合[1,9-11]。因此,巯基改性也成为一些重金属吸附与钝化材料的重要改性方法之一。如利用巯基改性的蒙脱石和坡缕石可以通过静电作用、离子交换及配位吸附等作用降低土壤中生物可利用镉的含量,有效实现土壤中镉的钝化[12-13];巯基改性的纤维素和煤矸石等材料可通过配位吸附等方式实现水中的汞和镉等重金属的高效去除[14-15]。巯基在生物体中广泛存在,主要来自蛋白或多肽上的半胱氨酸残基[9]。水中微生物(也包括生物来源的天然有机质)表面同样有巯基存在,尽管其浓度与羟基、羧基和氨基等基团相比相对较低,但其与重金属的结合效率往往远高于其他常见基团[9,11,16]。由于巯基环境浓度较低但结合重金属的能力较强,因此不同课题组的研究均发现吸附体系中重金属的浓度和负载量对细菌表面金属结合位点的类型有重要影响:当水中重金属浓度较高(mg/L级)或负载量较高时,细菌表面的羧基、磷酰基和氨基等浓度较高的基团是主要的金属结合位点;而当金属浓度较低(μg/L级)或负载量较低时,细菌表面的巯基对金属离子表现出很强的结合能力[7,16-18]。因此,在重金属浓度和负载量较低的自然水体中,水中存在的巯基可能会优先吸附结合重金属,成为影响重金属行为的重要影响因素。目前已有关于某些细菌表面巯基与重金属结合特征的研究工作[9,19-20],但对微藻的相关研究还鲜有报道,更未见对于水环境中的天然生物膜等复杂固相物质表面巯基与重金属结合情况的研究。由于自然水体中生物膜广泛存在,是水环境中微生物的主要存在形式之一,因此研究水体生物膜中巯基的存在及其对重金属的吸附结合特征为深入探究生物膜与金属相互作用的微观机理提供支撑,对于深入了解水环境中重金属的行为具有重要意义,也有助于全面认识自然水体中生物膜的作用和环境意义。
为区分巯基与其他基团对重金属的结合,现采用选择性掩蔽的方法[21-22]。其原理与用选择性萃取法来研究生物膜中特定组分对重金属的吸附规律类似[2,5],即利用与巯基有高选择性结合的试剂来掩蔽巯基,通过对比掩蔽前后生物膜对重金属吸附特征的差异,来确定被掩蔽巯基的吸附特征。
1 材料与方法
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1.1 生物膜的获取
本文研究取湖水在实验室中人工培养获取生物膜,湖水取自长春南湖。取30 L湖水于50 L的玻璃缸中,以清洗后的载玻片(48 mm × 75 mm × 1 mm,以聚丙烯架固定)为基质培养生物膜。培养在室温和自然光照下进行,培养期间定期调整聚丙烯架方位来保证各载玻片上生物膜尽可能均一。生物膜培养20 d左右,取生长均匀的载玻片,用微量矿物盐(minimal mineral salts, MMS)溶液[2,5]润洗后用于后续实验。生物膜的主要微生物种类用光学显微镜(XSJ-2,重庆光电仪器有限公司)观察确定。生物膜量用干重(103 ℃干燥至恒重)定量。生物膜中的酸可萃取金属量用15% HNO3溶液振荡萃取24 h后用火焰原子吸收分光光度计(AA6880,岛津制作所)测定。用总有机碳(total organic carbon, TOC)表征生物膜中有机质的含量(TOC-VCPH,配SSM-5000A型固体进样器,岛津制作所)。
1.2 qBBr与生物膜结合条件确定
选择5-(溴甲基)-N,N,N,2,6-五甲基-1,7-二氧代-1H,7H-吡唑并[2-A]吡唑-3-甲基溴化铵[monobromo(trimethylammonio)-bimane,qBBr,西格玛奥德里奇]为巯基特异性结合试剂来掩蔽生物膜中的巯基[21-22]。为保证掩蔽效果和对生物膜中微生物影响较小,先通过实验确定合适的qBBr与生物膜结合条件,主要包括:确定紫外可见分光光度法定量qBBr的方法,确定qBBr与生物膜结合的平衡时间,确定qBBr浓度对生物膜中微生物活性[以生物膜的叶绿素a含量、超氧化物歧化酶(superoxide dismutase, SOD)活性和丙二醛(malondialdehyde, MDA)含量为代表]的影响,确定qBBr浓度对其与生物膜结合的影响及饱和结合条件。
1.3 生物膜中巯基含量测定
利用酸碱电位滴定法(ZDJ-4A自动电位滴定仪,上海仪电科学仪器有限公司)分别测定qBBr屏蔽前后生物膜上氢离子结合位点的总浓度,二者差值即为生物膜中被掩蔽的巯基位点浓度(qBBr分子本身在实验pH范围内不与氢离子结合[21])。用1.2节中确定的掩蔽条件掩蔽巯基;利用4位点-非静电表面络合模型[11,21]拟合滴定数据,计算点位浓度。
1.4 生物膜对重金属的吸附实验
用硅胶刷刮下载玻片上的生物膜,在1.2节确定的掩蔽条件下用qBBr与生物膜反应。反应后的生物膜在4 000 r/min转速下离心5 min,重复两次,去除表面附着的qBBr。以Cu、Pb和Cd为常见重金属代表,每种金属分别用其硝酸盐和MMS溶液配制6种初始浓度(0.025、0.05、0.125、0.25、0.75、2.0 μmol/L)的吸附溶液,每个吸附体系加入qBBr掩蔽后的生物膜10.0 mg(干重,下同),吸附溶液体积40 mL。上述各体系中单位质量生物膜的最大理论金属负载量分别为0.1、0.2、0.5、1.0、3.0、8.0 μmol/g。每个金属的各浓度系列均分为4个不同的吸附pH(4.5、5.2、6.0和7.0),溶液pH用0.01 mol/L的HNO3和NaOH调节并控制。吸附在100 mL的具塞玻璃锥形瓶中进行,用水浴恒温振荡器于(25 ± 2) ℃下振荡吸附。吸附10 h后(根据预实验结果,吸附在10 h内平衡),将吸附溶液用0.45 μm混合纤维素滤膜抽滤,滤膜润洗后用25 mL、15% HNO3溶液振荡萃取24 h。用电感耦合等离子体质谱仪(7500C,安捷伦)测定吸附溶液中各金属的初始和平衡溶液,及HNO3萃取液中的各金属浓度。同法进行未掩蔽生物膜的上述吸附实验。生物膜吸附金属的热力学特征用Langmuir吸附等温线描述[2],即
$ \Gamma=\frac{\Gamma_{\max } K C}{1+K C}$
式(1)中:Γ 为生物膜对金属的平衡吸附量;Γmax为最大吸附量;K 为Langmuir平衡常数;C 为吸附平衡浓度。
2 结果与讨论
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2.1 qBBr与生物膜结合特征及生物膜表面巯基
根据显微镜观察,所培养生物膜含有大量微生物,包括小球藻属、舟形藻属、栅藻属的多种微藻、钟虫属的原生动物等,也含有菌胶团和矿物成分。生物膜干重为(17.8 ± 1.8) mg/片,生物膜的铁、锰和TOC的含量分别为(16.8 ± 0.23)、(14.3 ± 0.16)和(434.6 ± 15.9) mg/g。总体而言,本文研究所用生物膜与以往研究的生物膜[2,5,23]相似。
通过对qBBr溶液的全波段扫描,确定了其在243 nm处的特征吸收峰。该波长下,qBBr浓度与吸光度间线性关系良好,相关系数在0.999 0以上,可用于qBBr的定量测定。
从图1中qBBr与生物膜的结合过程可见,qBBr与生物膜在振荡条件下在前400 min内快速结合,600 min(10 h)后趋于平衡。由图2可见,在10 h内,各初始浓度的qBBr对生物膜中叶绿素a含量的影响很小;对生物膜SOD酶活性的影响在qBBr浓度在7.5 μmol/L以下时很小,在20 μmol/L及以上时有一定影响;膜中MDA含量随qBBr浓度升高而升高,但在qBBr浓度小于7.5 μmol/L时其影响相对较小。
从图3中不同浓度qBBr与生物膜作用10 h后的结合量看,结合量先随qBBr平衡浓度增加而较快增加,在平衡浓度达2.02 μmol/L后结合量随浓度进一步增加的量很小。在结合量随浓度较快增加阶段,主要应该是qBBr与巯基的大量结合;在缓慢增加阶段,主要应该是qBBr的表面吸附。qBBr平衡浓度为2.02 μmol/L时,对应的初始浓度为7.5 μmol/L,相应qBBr与生物膜质量(干重)比为10 μmol/g,生物膜对qBBr的平衡结合量为7.7 μmol/g。考虑过量qBBr在生物膜物质表面吸附对重金属吸附特征的可能影响,以及对微生物细胞膜通透性的可能影响[22],再综合考虑前述qBBr浓度对生物膜主要生理指标影响的情况,确定qBBr与生物膜的结合条件为初始浓度不超过7.5 μmol/L,结合体系的qBBr与生物膜干重比为10 μmol/g,振荡反应10 h。在此结合条件下,qBBr与生物膜结合充分且对生物膜代表性生理指标的影响较小或可忽略。结合前后的生物膜在光学显微镜下也未见有明显的差异:生物膜中微生物种类无明显改变,微藻细胞的外壁和结构无明显变化,细胞颜色无明显改变。
拟合滴定实验结果(图4),得到qBBr结合前后生物膜表观结合位点的表观解离常数pKa及其浓度如表1所示。由表1可知,生物膜中巯基结合位点主要为表观结合位点3,巯基浓度为(5.8 ± 0.6) μmol/g,约占生物膜表面总位点浓度的5.7%。本文研究所得生物膜巯基位点的表观pKa特征与相关细菌表面巯基的研究基本一致,但生物膜表面巯基浓度较细菌表面略低(细菌表面巯基一般在10到几十μmol/g)[15-18],与河水天然有机质(Suwannee河2R101N样品)的巯基浓度[(7.5 ± 0.4) μmol/g]较接近[24]。这一方面是由于生物膜中不仅有细菌、微藻、真菌等微生物成分,同时还存在较多的金属氧化物和其他矿物质等无机非生命成分,而非生命成分中巯基的浓度很低[25-27];另一方面是由于生物膜是在实际湖水中培养,湖水中存在的金属离子等成分也会在生物膜生长过程中结合到其表面巯基上,影响了巯基浓度的测定[2,28]。前面实验得出在qBBr与生物膜质量比为10 μmol/g时,生物膜对qBBr的平衡结合量为7.7 μmol/g,略大于算得的巯基浓度5.8 μmol/g。这说明生物膜表面对过量qBBr存在一定的吸附现象;也说明在前述所确定的qBBr与生物膜的结合条件下,生物膜表面巯基可被充分掩蔽。
2.2 生物膜中巯基对重金属的吸附特征
根据拟合结果,qBBr掩蔽前后的自然水体生物膜在不同pH下对Cu、Pb和Cd三种重金属的吸附均符合Langmuir吸附等温线。以Cu为例,吸附等温线如图5所示,相应拟合参数如表2所示。由表2可知,非线性拟合的R2 ≥ 0.996 5,表明实验数据较好地符合Langmuir吸附等温线。pH对吸附的影响与一般规律一致,即吸附量随pH的增加在一定范围内增加[1-2,29]。对比各pH下qBBr掩蔽前后生物膜的吸附等温线可见掩蔽均在一定程度上降低了Cu的吸附量,降低幅度与pH和平衡浓度有关。
以平衡浓度取2.0 μmol/L为例,代入根据表2以平衡浓度取2.0 μmol/L为例,代入根据表2参数所确定的各吸附等温式,可计算出各pH下qBBr掩蔽前后Cu吸附量的差值:不同pH下掩蔽降低了6.2%~8.0%的Cu吸附量。该比例略大于巯基占生物膜表面总位点的比例(5.7%),可能与巯基对重金属的优先吸附有关,也可能与吸附参数的拟合误差有关。此外,各pH下生物膜对重金属的吸附量均为Pb > Cu > Cd,与相关研究结果[2,25,30]一致。
虽然巯基较其他基团或吸附点位有更强的结合重金属能力,但由于其含量较低,在水中重金属浓度较高时其对吸附的贡献并不明显(图5)。在水中重金属浓度相对生物膜(及其吸附基团)浓度而言较低时,巯基与其他点位竞争时优先结合重金属的特性可能才会有较显著的表现。因此,为进一步探究重金属浓度较低时巯基结合重金属的情况,更清楚地展示掩蔽前后生物膜吸附重金属特征的差异,对前面的吸附等温线数据重新整理,以吸附体系中生物膜的最大理论负载量(即体系中投加的重金属全部被生物膜所吸附时的理论吸附量,简称“理论负载量”,μmol/g)为横坐标、被吸附重金属占吸附体系中重金属总量的比例(简称“吸附比例”)为纵坐标作图,仍以Cu为例,如图6所示。总体而言,在理论负载量较低时,由于吸附体系中金属总量较少而生物膜上的吸附点位充足,此时重金属的吸附比例较高。随理论负载量增加,体系中重金属浓度增大,金属间竞争生物膜表面点位的作用增大,使其吸附比例逐渐下降。相对而言,未掩蔽生物膜对重金属的吸附比例更高,且随理论负载量增加而下降的趋势更显著,特别是在理论负载量较低时。各pH下掩蔽前后生物膜吸附比例差异的规律类似:体系理论负载量小于1.0 μmol/g时,特别是小于0.5 μmol/g时,掩蔽前后生物膜对铜的吸附比例差异明显,且理论负载量越小,差异越大;当理论负载量大于等于1.0 μmol/g时,掩蔽前后生物膜的吸附比例差异较小且随理论负载量变化不大,与图5所反映出的差异规律类似。这证实了前述巯基在重金属浓度较低时能够优先结合重金属,会显著影响其吸附特征;而当重金属浓度较高,其对重金属吸附的贡献并不显著。
进一步计算各pH下掩蔽前后生物膜对重金属吸附比例的差值,并比较其随理论负载量变化的情况,如图7所示(包括Pb和Cd)。由图7可更直观地看出吸附比例的差值在理论负载量大于1.0 μmol/g时较小且基本不变,而在1.0 μmol/g以下时随负载量减小而增大,并在本实验设计的最低理论负载量,0.1 μmol/g,时达最大。以Cu为例,不同pH下吸附比例差值的最大值在14%~27%。从不同pH看,同一理论负载量下,pH较低时吸附比例差异往往较小而pH较高时则差异较大,这主要与不同pH下的吸附量大小相关。从不同金属看,各金属的规律基本一致,只是Pb的吸附比例差值在理论负载量较低时相对更大些。
上述结果表明自然水体生物膜表面存在一定量的巯基基团,且它们在重金属浓度较低的情况下对生物膜吸附重金属有重要贡献。针对一些细菌开展的研究已证实在低金属负载量的条件下,细菌细胞及胞外聚合物表面的巯基在它们对重金属的吸附中起重要作用,而在高金属负载量的条件下,吸附由非巯基基团/点位主导[7,9,17]。本文研究进一步证实对于在水环境中会显著影响污染物环境行为的典型天然复杂固相物质——自然水体生物膜,同样存在类似的现象和规律。只是由于生物膜中含有无机矿物成分,且在培养过程中其巯基可能已经结合了水体中的一些金属离子,使得生物膜中巯基的含量和它们对重金属吸附的贡献均略低于文献[15-18]的细菌的情况。而这也意味着生物膜中的巯基在实际水环境中的作用可能会更大。同时,本文研究的实验包括较宽的pH范围、涵盖几种常见重金属,这也说明了生物膜中巯基重要作用具有普遍性。此外,本文研究所用生物膜属典型的表层水自养生物膜,其中虽含各种细菌、真菌等微生物,但微藻是其最主要的微生物成分[2,23,26]。这也说明微藻表面同样存在巯基及巯基对重金属的吸附,而微藻的相关研究鲜有报道。
自然水体中生物膜广泛存在,是水环境中微生物的最主要存在形式之一;同时,实际天然水环境中溶解态重金属的浓度一般较低,往往在μg/L数量级或更低,相应的理论金属负载量也很低。在这种情况下,根据本文研究和前述相关研究,水环境中的生物膜和其他类似固相物质,特别是生物成分,上的巯基对这些固相物质吸附重金属的作用有重要贡献,对天然水中重金属的环境行为和效应有重要影响。而巯基的存在及其对金属离子的高亲和力也进一步保证了天然水中溶解态重金属的低浓度,有效降低了重金属的生态环境风险[9,20]。此外,在自然水体的底泥等厌氧环境中存在的硫离子(S2-)同样能高效结合重金属,但生成的金属硫化物在好氧环境下往往会迅速被氧化而重新释放出重金属[1,10]。不同于金属硫化物,由于巯基的相对稳定性,金属巯基化合物也有更好的稳定性,特别是微生物表面的低浓度金属巯基化合物可以在有氧的地表水环境中相对稳定地存在,这进一步体现了天然水环境中巯基的重要作用和自然水体生物膜的重要环境意义。
3 结论
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(1)本文研究建立了适用于自然水体生物膜中巯基掩蔽和研究巯基对重金属吸附贡献的方法,并利用该方法研究不同pH下生物膜中巯基对Cu、Pb和Cd三种重金属的吸附特征。
(2)自然水体生物膜表面存在一定量的巯基基团,尽管巯基的浓度较低,但仍由于其具有更强的重金属结合能力,使其在重金属浓度较低时对生物膜吸附重金属有重要贡献。
(3)生物膜中巯基的存在和对低浓度重金属的优先结合能力对天然水环境中重金属的环境行为和效应有重要影响。
基金
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  • 国家自然科学基金(42077343)
  • 吉林省自然科学基金(20230101092JC)
文献
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doi: 10.12404/j.issn.1671-1815.2405989
  • 接收时间：2024-08-09
  • 首发时间：2025-12-17
  • 出版时间：2025-06-28
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  • 收稿日期：2024-08-09
  • 修回日期：2025-04-01
基金
国家自然科学基金(42077343)
吉林省自然科学基金(20230101092JC)
作者信息
    1 吉林大学新能源与环境学院, 长春 130012
    2 国家管网集团建设项目管理分公司, 廊坊 065001
    3 吉林省长春生态环境监测中心, 长春 130012

通讯作者:

* 花修艺(1976—),男,汉族,江苏邳州人,博士,教授。研究方向:环境化学。E-mail:。
参考文献
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https://castjournals.cast.org.cn/joweb/kxjsygc/CN/10.12404/j.issn.1671-1815.2405989
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2种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
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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