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Article(id=1226460580693849043, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1226460576751206672, articleNumber=null, orderNo=null, doi=10.13343/j.cnki.wsxb.20250049, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1737043200000, receivedDateStr=2025-01-17, revisedDate=null, revisedDateStr=null, acceptedDate=1741104000000, acceptedDateStr=2025-03-05, onlineDate=1770340588972, onlineDateStr=2026-02-06, pubDate=1754236800000, pubDateStr=2025-08-04, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1770340588972, onlineIssueDateStr=2026-02-06, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1770340588972, creator=13701087609, updateTime=1770340588972, updator=13701087609, issue=Issue{id=1226460576751206672, tenantId=1146029695717560320, journalId=1192105938417971205, year='2025', volume='65', issue='8', pageStart='1', pageEnd='3812', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1770340588033, creator=13701087609, updateTime=1770363610188, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1226557138735117113, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1226460576751206672, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1226557138735117114, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1226460576751206672, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=3615, endPage=3629, ext={EN=ArticleExt(id=1226460581008421860, articleId=1226460580693849043, tenantId=1146029695717560320, journalId=1192105938417971205, language=EN, title=Screening and characterization of self-producing biosurfactant bacteria for dimethyl disulfide deodorization, columnId=1192149543992045670, journalTitle=Acta Microbiologica Sinica, columnName=Research Article, runingTitle=null, highlight=null, articleAbstract=

Microbial deodorization is an effective technology for treating odorous waste gases, in which microbial strains play a decisive role. [Objective] To screen the strains capable of degrading dimethyl disulfide (DMDS) and investigate their degradation efficiency and ability to produce surfactants under various conditions. [Methods] DMDS, a typical sulfur-containing odorous organic compound, was selected as the sole carbon source. A strain R1 capable of simultaneously producing biosurfactants and degrading DMDS was isolated from mangrove sludge. The strain was identified based on the physiological and biochemical characteristics analysis and 16S rRNA gene sequencing. The types of self-produced biosurfactants were determined using infrared spectroscopy and nuclear magnetic resonance spectroscopy analysis. [Results] Based on physiological and biochemical characteristics and 16S rRNA gene sequence, strain R1 was identified as Achromobacter sp. The strain was capable of degrading DMDS, with optimal degradation conditions of an initial DMDS concentration of 12.49 mg/L, a system temperature of 30 ℃, and an inoculum amount of 1.0 g/L, under which the DMDS degradation rate reached 70.74%. Emulsification experiments showed that strain R1 can use DMDS as a carbon source to produce biosurfactants, which were identified as glycolipids through nuclear magnetic resonance and infrared spectroscopy. [Conclusion] The main intermediate product in the biodegradation of DMDS is methyl mercaptan, and the transformation rate of sulfur to SO42- is 65.99%. Strain R1 exhibits impressive performance in degrading DMDS and producing biosurfactants.

, correspAuthors=Shungang WAN, authorNote=null, correspAuthorsNote=
*E-mail:
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微生物除臭是一种有效的恶臭废气处理技术，菌种对生物除臭效果具有决定性的影响。 【目的】 筛选可降解二甲基二硫醚(dimethyl disulfide, DMDS)的菌株，并探究其在不同条件下的降解效果及产表面活性剂的能力。 【方法】 以含硫恶臭有机物DMDS为唯一碳源，从红树林底泥中筛选可自产生物表面活性剂并能降解DMDS的菌株R1。利用生理生化特性分析和16S rRNA基因测序对菌株进行鉴定，利用红外光谱和核磁分析确定自产生物表面活性剂的种类。 【结果】 通过生理生化特性及16S rRNA基因序列分析鉴定，菌株R1属于木糖氧化无色杆菌(Achromobacter xylosoxidans)。研究表明，在DMDS初始浓度为12.49 mg/L、温度为30 ℃、接种量为1.0 g/L的条件下，DMDS的降解率可达70.74%。乳化实验表明，菌株R1能够利用DMDS作为碳源分泌生物表面活性剂，核磁和红外光谱分析鉴定表明，菌株R1产生的表面活性剂种类为糖脂类。 【结论】 DMDS降解的中间产物主要为甲硫醇，硫元素的转化率为65.99%。菌株R1表现出良好的降解能力和产表面活性剂性能。

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作者贡献声明

吴江丽：实验操作，数据收集和处理，论文撰写和修改；孙蕾：论文修改；袁丹：协助数据处理与分析；万顺刚：研究构思和设计，论文审阅和修改。

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Frontiers in Bioengineering and Biotechnology, 2022, 10: 794460., articleTitle=Rhamnolipids as green stabilizers of nZVI and application in the removal of nitrate from simulated groundwater, refAbstract=null), Reference(id=1226596311806427203, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226460580693849043, doi=null, pmid=null, pmcid=null, year=2025, volume=417, issue=null, pageStart=131833, pageEnd=null, url=null, language=null, rfNumber=[29], rfOrder=36, authorNames=ZHENG XY, LI YY, XU JC, ZHANG QX, ZHANG YX, journalName=Bioresource Technology, refType=null, unstructuredReference=ZHENG XY, LI YY, XU JC, ZHANG QX, ZHANG YX. Characterization of three novel dimethyl disulfide degrading bacteria and their potential degradation pathways[J]. Bioresource Technology, 2025, 417: 131833., articleTitle=Characterization of three novel dimethyl disulfide degrading bacteria and their potential degradation pathways, refAbstract=null), Reference(id=1226596311915479114, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226460580693849043, doi=null, pmid=null, pmcid=null, year=2022, volume=315, issue=null, pageStart=120469, pageEnd=null, url=null, language=null, rfNumber=[30], rfOrder=37, authorNames=ZHAO JK, GAO JL, JIN XY, YOU JP, FENG K, YE JX, CHEN JM, ZHANG SH, journalName=Environmental Pollution, refType=null, unstructuredReference=ZHAO JK, GAO JL, JIN XY, YOU JP, FENG K, YE JX, CHEN JM, ZHANG SH. Superior dimethyl disulfide degradation in a microbial fuel cell: extracellular electron transfer and hybrid metabolism pathways[J]. 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A-C: Colony morphology; D: Gram staining; E-F: Scanning electron microscopy image (E: 10 000×; F: 50 000×)., figureFileSmall=HMPT8u5UoRcVdWI2P6RWEQ==, figureFileBig=0Z2rjAihCTuFvDQVYA1D4A==, tableContent=null), ArticleFig(id=1226596301463274116, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226460580693849043, language=CN, label=图1, caption=菌株R1的形态特征。A-C：菌落形态图；D：革兰氏染色图；E-F：扫描电镜图(E为放大10 000倍，F为放大50 000倍)。, figureFileSmall=HMPT8u5UoRcVdWI2P6RWEQ==, figureFileBig=0Z2rjAihCTuFvDQVYA1D4A==, tableContent=null), ArticleFig(id=1226596301618463372, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226460580693849043, language=EN, label=Figure 2, caption=Phylogenetic tree of strain R1 based on 16S rRNA gene sequence. GenBank accession numbers are provided in parentheses. The Bootstrap value based on 1 000 repeated calculations are shown at the branch point., figureFileSmall=LO1/jh8k4TkJ6VKWsyh2pg==, figureFileBig=RT0Zp6ixJ0bbnxpEJ8rdWw==, tableContent=null), ArticleFig(id=1226596301798818450, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226460580693849043, language=CN, label=图2, caption=菌株R1基于16S rRNA基因序列构建的系统发育树。括号内为GenBank登录号；分支点显示了基于 1 000次重复计算得出的Bootstrap值。, figureFileSmall=LO1/jh8k4TkJ6VKWsyh2pg==, figureFileBig=RT0Zp6ixJ0bbnxpEJ8rdWw==, tableContent=null), ArticleFig(id=1226596301878510234, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226460580693849043, language=EN, label=Figure 3, caption=Effect of initial DMDS concentration on biodegradation performance. A: DMDS concentration in gas phase; B: DMDS concentration in liquid phase; C: Degradation rate; D: DMDS mineralization rate. Degradation conditions: Inoculation amount 1.0 g/L, 35 °C., figureFileSmall=lvq22Tl5ioVbtdDmf/j+2A==, figureFileBig=zFmg/1xh2InGNcikRHaJ2g==, tableContent=null), ArticleFig(id=1226596301995950754, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226460580693849043, language=CN, label=图3, caption=DMDS初始浓度对降解效果的影响。A：气相中DMDS浓度；B：液相中DMDS浓度；C：DMDS降解率；D：DMDS矿化率。降解条件：接种量1.0 g/L，35 ℃。, figureFileSmall=lvq22Tl5ioVbtdDmf/j+2A==, figureFileBig=zFmg/1xh2InGNcikRHaJ2g==, tableContent=null), ArticleFig(id=1226596302100808363, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226460580693849043, language=EN, label=Figure 4, caption=Effect of inoculation amount of strain R1 on DMDS biodegradation performance. A: DMDS concentration in gas phase; B: DMDS concentration in liquid phase; C: Degradation rate; D: DMDS mineralization rate. Degradation conditions: DMDS 12.49 mg/L, 35 °C., figureFileSmall=tXfZ/MyYRoJs6TtaFDXU9g==, figureFileBig=9EQE4kikwbtsbIi2W4boLw==, tableContent=null), ArticleFig(id=1226596302230831794, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226460580693849043, language=CN, label=图4, caption=菌株R1添加量对DMDS降解效果的影响。A：气相中DMDS浓度；B：液相中DMDS浓度；C：DMDS降解率；D：DMDS矿化率。降解条件：DMDS 12.49 mg/L，35 ℃。, figureFileSmall=tXfZ/MyYRoJs6TtaFDXU9g==, figureFileBig=9EQE4kikwbtsbIi2W4boLw==, tableContent=null), ArticleFig(id=1226596302335689404, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226460580693849043, language=EN, label=Figure 5, caption=Effect of temperature on DMDS biodegradation performance. A: DMDS concentration in gas phase; B: DMDS concentration in liquid phase; C: Degradation rate; D: DMDS mineralization rate. Degradation conditions: DMDS 12.49 mg/L, inoculation amount 1.0 g/L., figureFileSmall=JOboQHEWo0qRVG9bH0FRtA==, figureFileBig=Lgo56YNk/+yVxn7Mwy+/Dg==, tableContent=null), ArticleFig(id=1226596302482490054, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226460580693849043, language=CN, label=图5, caption=体系温度对DMDS降解效果的影响。A：气相中DMDS浓度；B：液相中DMDS浓度；C：DMDS降解率；D：DMDS矿化率。降解条件：DMDS 12.49 mg/L，接种量1.0 g/L。, figureFileSmall=JOboQHEWo0qRVG9bH0FRtA==, figureFileBig=Lgo56YNk/+yVxn7Mwy+/Dg==, tableContent=null), ArticleFig(id=1226596302595736270, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226460580693849043, language=EN, label=Figure 6, caption=Effect of DMDS concentration on emulsifying ability of biosurfactant produced by strain R1. A: Degradation time 5 days; B: Complete degradation., figureFileSmall=Ggi0Ek3Y7oA7tPz/IvYlTQ==, figureFileBig=NAG5fPyO7A7gxOh/41YFuw==, tableContent=null), ArticleFig(id=1226596302717371096, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226460580693849043, language=CN, label=图6, caption=DMDS浓度对菌株R1自产生物表面活性剂能力的影响。A：降解时间第5天；B：完全降解。, figureFileSmall=Ggi0Ek3Y7oA7tPz/IvYlTQ==, figureFileBig=NAG5fPyO7A7gxOh/41YFuw==, tableContent=null), ArticleFig(id=1226596302813840094, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226460580693849043, language=EN, label=Figure 7, caption=FT-IR spectra and nuclear magnetic resonance spectroscopy. A: FT-IR spectra of biosurfactants secreted by strain R1; B: FT-IR spectra of rhamnolipid; C: Nuclear magnetic resonance spectroscopy (1H NMR) of biosurfactants secreted by strain R1; D: Nuclear magnetic resonance spectroscopy of rhamnolipid., figureFileSmall=5FLvWvKgeo1MuywfT91CXw==, figureFileBig=lRI2pqSjeszHQHEYN2XxJQ==, tableContent=null), ArticleFig(id=1226596304193766118, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226460580693849043, language=CN, label=图7, caption=FT-IR光谱和核磁共振波谱。A：菌株R1分泌的表面活性剂的FT-IR光谱；B：鼠李糖脂的FT-IR光谱；C：菌株R1分泌的生物表面活性剂的核磁共振波谱；D：鼠李糖脂的核磁共振波谱。, figureFileSmall=5FLvWvKgeo1MuywfT91CXw==, figureFileBig=lRI2pqSjeszHQHEYN2XxJQ==, tableContent=null), ArticleFig(id=1226596304311206636, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226460580693849043, language=EN, label=Figure 8, caption=Mass spectra of DMDS and intermediates. A: Dimethyl disulfide; B: Bis (methylthio) methane; C: Methyl mercaptan., figureFileSmall=EZcXmq+buBKmiY03c5zaMg==, figureFileBig=++Lfzp/RPJM/HhOgoWNDWw==, tableContent=null), ArticleFig(id=1226596304432841463, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226460580693849043, language=CN, label=图8, caption=DMDS和中间产物的质谱图。A：二甲基二硫醚；B：二甲硫基甲烷；C：甲硫醇。, figureFileSmall=EZcXmq+buBKmiY03c5zaMg==, figureFileBig=++Lfzp/RPJM/HhOgoWNDWw==, tableContent=null), ArticleFig(id=1226596304550281981, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226460580693849043, language=EN, label=Figure 9, caption=Possible degradation pathways of DMDS., figureFileSmall=KYf4SLTQMZ+73zRmNYYKiw==, figureFileBig=974gbYqh6cRK2mCjJ2j3aA==, tableContent=null), ArticleFig(id=1226596304722248457, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226460580693849043, language=CN, label=图9, caption=生物降解DMDS的可能途径。, figureFileSmall=KYf4SLTQMZ+73zRmNYYKiw==, figureFileBig=974gbYqh6cRK2mCjJ2j3aA==, tableContent=null), ArticleFig(id=1226596304885826324, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226460580693849043, language=EN, label=Figure 10, caption=The relationship between the changes in DMDS concentration and the accumulation of SO42- concentration., figureFileSmall=C1Ap9acdshFcaJhl6ZfQWQ==, figureFileBig=PZXeB57Ydj+kuAU19ogQOQ==, tableContent=null), ArticleFig(id=1226596304982295322, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226460580693849043, language=CN, label=图10, caption=DMDS浓度变化与SO42-累积量的关系。, figureFileSmall=C1Ap9acdshFcaJhl6ZfQWQ==, figureFileBig=PZXeB57Ydj+kuAU19ogQOQ==, tableContent=null), ArticleFig(id=1226596305124901666, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226460580693849043, language=EN, label=Table 1, caption=

Physiological and biochemical characteristics

, figureFileSmall=null, figureFileBig=null, tableContent=
Physiological and biochemical experimentsResults
Enzyme+
Oxidase+
Anaerobic growth-
Nitrate reduction+
Utilization of citrate+
Urease-
Gelatin liquefaction-
Hemolysis-
Lysine decarboxylase-
Arginine dihydrolase-
Glucose fermentation+
Galactose fermentation+
Mannose fermentation+
), ArticleFig(id=1226596305250730794, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1226460580693849043, language=CN, label=表1, caption=

生理生化特性

, figureFileSmall=null, figureFileBig=null, tableContent=
Physiological and biochemical experimentsResults
Enzyme+
Oxidase+
Anaerobic growth-
Nitrate reduction+
Utilization of citrate+
Urease-
Gelatin liquefaction-
Hemolysis-
Lysine decarboxylase-
Arginine dihydrolase-
Glucose fermentation+
Galactose fermentation+
Mannose fermentation+
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自产生物表面活性剂的二甲基二硫醚除臭功能菌筛选及其性能分析
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吴江丽 1 , 孙蕾 2 , 袁丹 2 , 万顺刚 2, *
微生物学报 | 研究报告 2025,65(8): 3615-3629
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微生物学报 | 研究报告 2025, 65(8): 3615-3629
自产生物表面活性剂的二甲基二硫醚除臭功能菌筛选及其性能分析
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吴江丽1, 孙蕾2, 袁丹2, 万顺刚2, *
作者信息
  • 1.海南大学 生态学院，海南 海口
  • 2.海南大学 化学化工学院，海南 海口
Screening and characterization of self-producing biosurfactant bacteria for dimethyl disulfide deodorization
Jiangli WU1, Lei SUN2, Dan YUAN2, Shungang WAN2, *
Affiliations
  • 1.School of Ecology, Hainan University, Haikou, Hainan, China
  • 2.School of Chemistry and Chemical Engineering, Hainan University, Haikou, Hainan, China
出版时间: 2025-08-04 doi: 10.13343/j.cnki.wsxb.20250049
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摘要
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微生物除臭是一种有效的恶臭废气处理技术，菌种对生物除臭效果具有决定性的影响。 【目的】 筛选可降解二甲基二硫醚(dimethyl disulfide, DMDS)的菌株，并探究其在不同条件下的降解效果及产表面活性剂的能力。 【方法】 以含硫恶臭有机物DMDS为唯一碳源，从红树林底泥中筛选可自产生物表面活性剂并能降解DMDS的菌株R1。利用生理生化特性分析和16S rRNA基因测序对菌株进行鉴定，利用红外光谱和核磁分析确定自产生物表面活性剂的种类。 【结果】 通过生理生化特性及16S rRNA基因序列分析鉴定，菌株R1属于木糖氧化无色杆菌(Achromobacter xylosoxidans)。研究表明，在DMDS初始浓度为12.49 mg/L、温度为30 ℃、接种量为1.0 g/L的条件下，DMDS的降解率可达70.74%。乳化实验表明，菌株R1能够利用DMDS作为碳源分泌生物表面活性剂，核磁和红外光谱分析鉴定表明，菌株R1产生的表面活性剂种类为糖脂类。 【结论】 DMDS降解的中间产物主要为甲硫醇，硫元素的转化率为65.99%。菌株R1表现出良好的降解能力和产表面活性剂性能。

二甲基二硫醚  /  生物降解  /  分离鉴定  /  生物表面活性剂  /  除臭

Microbial deodorization is an effective technology for treating odorous waste gases, in which microbial strains play a decisive role. [Objective] To screen the strains capable of degrading dimethyl disulfide (DMDS) and investigate their degradation efficiency and ability to produce surfactants under various conditions. [Methods] DMDS, a typical sulfur-containing odorous organic compound, was selected as the sole carbon source. A strain R1 capable of simultaneously producing biosurfactants and degrading DMDS was isolated from mangrove sludge. The strain was identified based on the physiological and biochemical characteristics analysis and 16S rRNA gene sequencing. The types of self-produced biosurfactants were determined using infrared spectroscopy and nuclear magnetic resonance spectroscopy analysis. [Results] Based on physiological and biochemical characteristics and 16S rRNA gene sequence, strain R1 was identified as Achromobacter sp. The strain was capable of degrading DMDS, with optimal degradation conditions of an initial DMDS concentration of 12.49 mg/L, a system temperature of 30 ℃, and an inoculum amount of 1.0 g/L, under which the DMDS degradation rate reached 70.74%. Emulsification experiments showed that strain R1 can use DMDS as a carbon source to produce biosurfactants, which were identified as glycolipids through nuclear magnetic resonance and infrared spectroscopy. [Conclusion] The main intermediate product in the biodegradation of DMDS is methyl mercaptan, and the transformation rate of sulfur to SO42- is 65.99%. Strain R1 exhibits impressive performance in degrading DMDS and producing biosurfactants.

dimethyl disulfide  /  biodegradation  /  isolation and identification  /  biosurfactant  /  deodorization
吴江丽, 孙蕾, 袁丹, 万顺刚. 自产生物表面活性剂的二甲基二硫醚除臭功能菌筛选及其性能分析. 微生物学报, 2025 , 65 (8) : 3615 -3629 . DOI: 10.13343/j.cnki.wsxb.20250049
Jiangli WU, Lei SUN, Dan YUAN, Shungang WAN. Screening and characterization of self-producing biosurfactant bacteria for dimethyl disulfide deodorization[J]. Acta Microbiologica Sinica, 2025 , 65 (8) : 3615 -3629 . DOI: 10.13343/j.cnki.wsxb.20250049
正文
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挥发性有机硫化合物(volatile organic sulfur compounds, VOSCs)具有非常低的嗅阈值和高毒性，极易引起空气污染，是典型的恶臭污染物之一[1]。VOSCs主要包括二甲基三硫醚、甲硫醇、二甲基硫醚和二甲基二硫醚(dimethyl disulfide, DMDS)等[2]，在各种污染源排放的VOSCs中DMDS的检出频率和浓度均高于其他VOSCs。崔健等[3]对无锡市5处典型重度黑臭水体连续5个月采样分析，结果表明DMDS是检测到的唯一VOSCs，检出频率为88%，最高浓度为420.85 ng/L。丁家正等[4]总结了城镇污水处理厂各处理单元VOSCs的排放特征，结果发现均有DMDS排放，并且在多数单元检测到的DMDS浓度均高于其他VOSCs。因此，采取有效的处理技术去除以DMDS为代表的VOSCs具有重要的研究意义。目前，VOSCs的去除方法主要有物理、化学和生物等方法。其中，物理法主要包括吸附、冷凝和吸收；化学法主要有光催化、焚烧和等离子体技术等[5]。与物理和化学方法相比，生物法是利用微生物将VOSCs作为碳源和能量，通过自身代谢降解VOSCs的方法[6]，生物法具有节能环保的潜在优势，特别适用于处理废气中低浓度的VOSCs。因此，微生物法可以有效解决其他VOSCs处理技术存在的降解效率低以及副产物残留的问题[7]。然而，VOSCs均为疏水性有机物，其难溶于水的特性导致其生物利用性差，已成为生物降解疏水性VOSCs的瓶颈问题。
现有研究表明，在生物降解疏水性有机物的过程中添加表面活性剂，通过降低界面表面张力和形成胶束，强化气液两相传质，促进疏水性有机物在液相中的溶解性，被认为是一种行之有效的生物降解强化策略，具有极大的应用潜力[8]。Dewidar和Sorial[9]在接种真菌的生物滴滤池中添加生物表面活性剂鼠李糖脂，降低了表面张力并形成胶束，提高了甲苯的扩散速率，从而增强了气相甲苯的去除效果。此外，鼠李糖脂在适当浓度下有利于微生物生长。Lamprea-Pineda等[10]在生物过滤反应器中添加表面活性剂皂苷，强化了2种疏水性VOCs (环己烷和己烷混合物)的生物降解效果，发现环己烷和己烷在生物过滤器中的停留时间缩短，去除率提高了26%。这表明生物表面活性剂在增效去除疏水性VOCs方面具有很好的潜力。与外源添加的人工合成化学表面活性剂可能带来的毒性相比，利用微生物在降解疏水性VOSCs的同时自产生物表面活性剂，强化降解过程中疏水性VOSCs的气液两相传质，具有更低的生物毒性和更好的环境兼容性。尽管目前已有相关研究报道了VOSCs降解菌种的筛选，但对于以VOSCs为唯一营养源，兼具降解和自产生物表面活性剂能力的菌种筛选，相关研究仍较为缺乏。
本研究针对生物降解疏水性VOSCs过程中存在的问题，以典型的含硫恶臭污染物DMDS为目标污染物，以海南省海口市东寨港红树林湿地的底泥为筛选菌源，从中筛选兼具降解DMDS能力以及分泌生物表面活性剂能力的功能性菌株。通过16S rRNA基因测序和生理生化性质鉴定菌株的属性，并采用乳化实验和核磁等分析手段明确了所产生物表面活性剂的种类。结合气相色谱质谱(GC/MS)分析降解过程中生成的中间产物，提出可能的生物降解机理。
1 材料与方法
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1.1 材料
1.1.1 菌种来源
海南省海口市东寨港红树林湿地的底泥作为筛选DMDS降解菌株的菌源。
1.1.2 培养基
LB培养基(g/L)：蛋白胨10.0，牛肉膏3.0，氯化钠5.0。亚甲基蓝培养基(g/L)：蛋白胨5.000，酵母提取物0.200，琼脂18.000，葡萄糖20.000，牛肉提取物1.000，亚甲基蓝0.005，十六烷基三甲基溴化铵0.200。血琼脂平板培养基购自广东环凯生物科技有限公司。无机盐培养基MSM(g/L)：K2HPO4·3H2O 1.20，MgSO4·7H2O 0.20，FeSO4·7H2O 0.01，KH2PO4 1.20，NH4Cl 0.40，微量元素液1.00 mL。微量元素液组成(g/L)：CaCl2 0.200，H3BO3 0.006，CoCl2·6H2O 0.090，CuSO4·5H2O 0.100，MnSO4·4H2O 0.200，Na2MoO4·2H2O 0.120，ZnSO4·7H2O 0.200。
1.2 兼具DMDS降解和自产生物表面活性剂菌株的筛选
将10 mL新鲜红树林底泥样品接种到含90 mL MSM和20.82 mg/L DMDS的650 mL顶空瓶中，35 ℃、100 r/min培养5 d进行DMDS降解菌的富集和筛选。将初步筛选的菌种培养物接种到血琼脂平板上，在35 ℃下进一步培养2-3 d，通过观察平板上菌落的均匀性和差异，挑取单个菌落进一步涂布在亚甲基蓝平板上，在35 ℃下培养2-3 d分离出能在血琼脂平板和亚甲基蓝平板上生长的细菌菌株。最后，将得到的菌落重新接种到LB培养基中，35 ℃、100 r/min培养24 h。培养完成后，将细胞悬液在常温10 000×g离心2 min，取湿基固体菌株接种到100 mL含有12.49 mg/L DMDS的MSM液体培养基中，35 ℃、100 r/min继续驯化和筛选。通过在血琼脂平板、亚甲基蓝平板、LB培养基和MSM液体培养基中进行多轮培养驯化和分离，最终得到能够以DMDS为唯一营养源进行生长的菌种，命名为菌株R1。
1.3 菌种鉴定
菌株R1的生理生化性质鉴定和分子生物学检测由广东省微生物分析检测中心完成。菌种同源性分析：通过BLAST程序，将菌株R1的16S rRNA基因序列与GenBank中已登录的微生物菌种序列进行同源性比较。利用开源软件MEGA 11，采用邻接法构建系统发育树，Bootstrap值为1 000次重复。
1.4 生物降解DMDS性能研究
从平板上挑取已经分离好的菌种置于灭菌的LB培养基中培养24 h。培养液在10 000×g下离心2 min，沉淀物用浓度为8%的灭菌无机盐溶液洗涤后再离心，重复洗涤离心3次，去除LB培养基并获得菌体。在顶空瓶中依次加入100 mL灭菌处理的MSM培养基溶液和适量菌体配制成菌悬液，并用带聚四氟乙烯垫片的顶空瓶盖密封。利用精密液体取样针准确移取适量液体DMDS，并快速注入到顶空反应器的内壁上，液体迅速挥发成DMDS气体。采用公式(1)计算气相中DMDS的初始浓度。
C0=ρV1V0
式中：C0为DMDS的初始浓度(mg/L)；ρ为DMDS的密度(1.062 5×106 mg/L)；V1为注入液体DMDS的体积(L)；V0为反应器内的气相体积(0.550 L)。
通过改变注入液体DMDS的体积，调控顶空瓶内气态DMDS的初始浓度分别为4.16、8.33、12.49、16.66和20.82 mg/L，研究DMDS初始浓度对菌种降解性能的影响。同时，按照相同的步骤，将含有100 mL MSM培养基和12.49 mg/L气态DMDS的顶空瓶分别在不同温度(25、30、35和40 ℃)和不同湿基生物量(0.6、1.0、1.4、1.8和2.2 g/L)下进行培养，定时取样分析气相和液相中DMDS的浓度，并计算DMDS的降解率。
气相中DMDS的检测方法：采用500 μL气密针(Agilent公司)，抽取顶空瓶上方的气体200 μL，注入配备FID检测器及HP-5 MS毛细管柱的气相色谱仪(上海天美科学仪器有限公司)进行检测分析。检测条件为：进样器温度180 ℃，色谱柱温度60 ℃，检测器温度220 ℃。液相中DMDS的检测方法：从顶空瓶内抽取0.5 mL液体培养基，注入含有1 mL二氯甲烷的色谱进样瓶进行原位萃取，取1 μL萃取液注入配有HP-5 MS毛细管柱的气相色谱质谱联用仪(Agilent Technologies公司)进行检测分析。检测条件：色谱柱温度100 ℃，质谱检测器温度230 ℃，进样器温度200 ℃。
DMDS的生物降解率计算如公式(2)所示。
降解率=C0-CC0×100%  
式中：C0和C分别为DMDS的初始浓度和剩余浓度(mg/L)。
1.5 乳化能力的测定
以花生油为模拟油相，以溶于液体培养基中的自产表面活性剂为乳化剂，通过计算乳化指数(E24)评估菌种自产表面活性剂的能力[11]。具体操作方法为：菌株R1在含有气态DMDS (12.49、16.66和20.82 mg/L)作为唯一碳源的100 mL MSM培养基中进行培养。待DMDS完全降解后10 000×g离心2 min，获得含表面活性剂的无细胞上清液。取3 mL花生油加入试管中，加入等量的无细胞上清液于同一试管，振荡1 min。制备的乳液在常温下沉降24 h，观察并记录乳化的高度，乳化指数计算如公式(3)所示[12]。
乳化指数E24=HeHT×100%
式中：He表示乳化层的高度(cm)；HT表示溶液的总高度(cm)。
1.6 生物表面活性剂的种类鉴定
将1 mL DMDS加入含有菌株R1溶液的100 mL MSM培养基顶空瓶中，并将顶空瓶在30 ℃、100 r/min下培养14 d。用6 mol/L盐酸将菌液离心后所得无细胞上清液的pH值调整到2.0，4 ℃静置24 h。利用等体积的乙酸乙酯萃取沉积物，重复3次，并在40 ℃的旋转蒸发仪中蒸发除去有机相中的乙酸乙酯以提取纯生物表面活性剂。收集有机相约1 mL，使用核磁共振波谱仪(NMR，Bruker BioSpin AG公司)分析生物表面活性剂的组成，频率为400 MHz，溶剂为氘代甲醇(CD3OD)。利用傅里叶变换红外光谱(FT-IR，Bruker公司)分析生物表面活性剂的化学基团。
1.7 降解中间产物和代谢终产物的测定
将菌株R1接种于LB培养基中，30 ℃、100 r/min培养24 h。培养液在常温下10 000×g离心2 min后，用MSM培养基洗净细菌菌体，并将其转移到含有100 mL MSM培养基的650 mL顶空瓶中，注入1 mL DMDS，并在摇床转速100 r/min、温度30 ℃、pH 7.0条件下培养10 d。10 d后10 000×g离心2 min收集无细胞上清液，用等体积的乙酸乙酯进行提取，再用旋转蒸发仪将有机相浓缩至1 mL。通过气相色谱质谱法测定DMDS的降解产物。气相色谱质谱仪的柱箱温度从40 ℃以5 ℃/min的速率逐渐升温至100 ℃，进样器温度设置为200 ℃，检测器温度设置为230 ℃。所有中间产物的化学结构均通过质谱分析获得。
通过离子色谱对S元素降解终产物SO42-进行定量分析测定。将菌株R1在LB培养基中富集培养24 h，10 000×g离心2 min后用无机盐溶液洗净菌体同样条件再次离心后将其转移到含有100 mL MSM培养基的顶空瓶中，DMDS浓度设置为16.66 mg/L，并在30 ℃、pH 7.0条件下分别培养3、5、10 d。以10 000×g离心2 min收集无细胞上清液5 mL，依次经过C18柱和0.22 μm针筒式滤膜过滤器过滤处理后，收集1 mL滤液，注入离子色谱(Dionex公司)进行浓度检测。分析条件：淋洗液为15 mmol/L KOH，流速为0.6 mL/min，色谱柱为SH-AP-1。
采用矿化率评估菌株R1生物降解DMDS生成CO2的能力，计算如公式(4)所示。气相中CO2浓度的检测方法：采用气密针(Agilent公司)抽取顶空瓶上方的气体200 μL，注入配备镍转化炉的气相色谱仪(浙江福立分析仪器有限公司)分析气相中CO2浓度。检测条件为：进样器温度180 ℃，色谱柱温度80 ℃，FID检测器温度200 ℃，镍转化炉温度350 ℃，色谱柱为HP-5 MS毛细管柱(30 m×0.32 mm×0.25 μm)。
矿化率=CACT×100%   
式中：CA为菌株R1生物降解DMDS实际生成CO2的量(mg/L)；CT为DMDS完全降解后理论上生成的CO2量(mg/L)。
2 结果与分析
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2.1 菌株的鉴定和表征
菌株R1在营养琼脂平板上恒温培养2 d后(图1A)，菌落呈圆形，凸起，表面光滑，边缘整齐。菌株R1可以在血琼脂平板(图1B)和亚甲基蓝平板(图1C)上生长，并表现出表面湿润、光滑、透明、微隆起、边缘较整齐、胶状、黏性强的特征。菌株R1革兰氏染色结果为红色(图1D)，说明其为革兰氏阴性菌。采用扫描电子显微镜进行形态学观察，发现菌株R1呈短杆状、表面光滑(图1E、1F)。结合表1所示的生理和生化特性，菌株R1是一株革兰氏阴性兼性厌氧菌，糖利用试验均呈阳性。
在GenBank中将菌株R1的16S rRNA基因测序序列与已知菌种序列比对，并构建系统发育树，结果如图2所示。菌株R1与木糖氧化无色杆菌(Achromobacter xylosoxidans)具有最高的相似性，相似度达100%，说明菌株R1属于无色杆菌属(Achromobacter sp.)，该菌株R1在广东省微生物菌种保藏中心的保藏号为3GDMCC. NO 65642。
2.2 DMDS降解条件优化
2.2.1 DMDS初始浓度的影响
DMDS初始浓度对菌株R1降解效果的影响如图3所示。随着DMDS初始浓度从4.16 mg/L增加到20.82 mg/L，气相(图3A)和液相(图3B)中DMDS的浓度逐渐增加，总的DMDS降解率逐渐下降(图3C)，而DMDS的矿化率提高(图3D)，表明初始浓度显著影响微生物对DMDS的利用效率。生物降解效率在最初的48 h内迅速提高，随后逐渐趋于平稳，这与Liang等[13]利用微生物降解DMDS的研究结果一致。液相中DMDS的残留量极少，即使在DMDS浓度为20.82 mg/L时，总降解率也达到了60.33%。这表明菌株R1本身分泌的表面活性剂起到了一定作用，使疏水性的DMDS能够更好地溶解于水中，从而被细菌利用。
2.2.2 接种量的影响
菌株R1的添加量对DMDS去除效果的影响如图4所示。随着菌株R1添加量的增加，对气相中DMDS的浓度变化不明显(图4A)，而液相中DMDS的浓度呈现先降低后增加的变化趋势(图4B)。菌种添加量对DMDS生物降解率的影响结果(图4C)表明，过高的菌种添加量并不会明显促进DMDS的生物降解。菌株R1添加量对DMDS矿化率的影响结果(图4D)表明，当DMDS作为唯一碳源时，菌种生物量越大，DMDS的矿化效果越明显。当菌株R1投加量过低时，培养液中菌种数量较少，导致菌种整体增殖缓慢。当菌种投加量过高时，菌种会加快DMDS的分解代谢并促进代谢产物的累积，导致菌种后期生长所需的营养缺乏。在这种情况下，局部环境中会产生菌株之间的竞争作用，从而抑制生物活性并影响DMDS的生物降解率。这一结果与其他研究者关于菌种添加量对不同污染物生物降解效果的研究结果一致[14-15]。因此，后续研究中采用1.0 g/L的菌种接种量。
2.2.3 温度的影响
在DMDS浓度为12.49 mg/L、pH 7.0和生物量1.0 g/L的条件下，研究了体系温度变化对菌株R1降解DMDS效率的影响(图5)。当温度在25-35 ℃范围内时，温度升高对气相和液相中DMDS浓度的变化影响显著，对降解率的增加具有一定的促进作用，总降解率分别为51.72%、70.74%和70.31%，说明温度的提高有利于微生物活性的增加。然而，当温度升高至40 ℃时，DMDS降解率降低至51.08%。这表明过高或过低的温度都会对微生物的生长和活性产生抑制作用，进而抑制生物降解活性。从上述结果可以看出，在pH 7.0、100 r/min、生物量1.0 g/L和DMDS浓度12.49 mg/L的条件下，菌株R1去除DMDS的最佳温度为30 ℃。这主要是因为微生物对温度较为敏感，在较低或较高的温度下对DMDS的去除能力均较差[16]。
2.3 表面活性剂的鉴定
采用乳化指数对菌株R1降解DMDS过程中产表面活性剂的能力进行了分析，结果如图6所示。在降解时间达到5 d时，DMDS浓度为12.49、16.66和20.82 mg/L时的乳化能力分别为36.58%、34.06%和31.74%。DMDS完全降解后，乳化能力有一定提高，分别为38.37%、36.84%和34.29%。在DMDS的生物降解过程中，乳化指数在DMDS浓度为12.49 mg/L时达到最佳。DMDS降解完成后，乳化指数达到最大值，说明在DMDS降解过程中，生物表面活性剂的含量逐渐增加。因此在生物利用DMDS的过程中，适当浓度的DMDS促进了菌株R1表面活性剂的产生，同时提高了DMDS的生物利用率，从而加快了DMDS的降解速率。
菌株R1自产生物表面活性剂的FT-IR和1H NMR表征结果如图7所示。图7A为自产生物表面活性剂样品的FT-IR光谱图。-OH的振动特征峰出现在3 444 cm-1和3 461 cm-1，证实了羟基的存在。2 848-2 995 cm-1处的双带归属于脂肪族群的C-H拉伸振动。1 756-1 758 cm-1处的特征峰对应酯基和羧酸基团的C=O拉伸键。1 245-1 379 cm-1之间的波段是碳水化合物C-H和O-H振动的典型特征，通常为鼠李糖单元[17]。1 054-1 056 cm-1的峰是最重要的特征峰，它们直接指示了糖苷键中的C-O和C-O-C的存在。同时，将自产生物表面活性剂的FT-IR谱图与典型生物表面活性剂(图7B)进行对比分析，结果表明菌株R1利用DMDS自产的生物表面活性剂与鼠李糖脂的表征结果高度相似，证明该菌株产生的生物表面活性剂为糖脂类。
此外，根据1H NMR波谱进一步分析了菌株R1自产的生物表面活性剂的类型，并将其与鼠李糖脂的1H NMR光谱进行比较，结果如图7C和图7D所示。化学位移在3.3 ppm的峰是由CD3OD溶剂引起的。1H NMR谱图显示，信号分别归属于脂质(脂肪酸)和糖(碳水化合物)部分[18]。脂肪酸官能团-(CH2)n和-C-OH基团分别位于化学位移1.22-1.26 ppm处和2.01-2.17 ppm范围[19]。关键的指示峰出现在化学位移3.5-5.2 ppm处，表明存在糖苷键，信号显示存在糖分子的糖苷键[20]。因此，可进一步判断菌株R1自产的生物表面活性剂为糖脂类。
2.4 DMDS的降解机理
采用GC/MS测定DMDS降解过程中生成的中间产物，结果发现菌株R1降解DMDS过程中共检出2个中间产物：二甲硫基甲烷(S1)和甲硫醇(S2) (图8)。根据检测到的中间产物种类并结合相关文献，推测DMDS的可能生物降解途径如图9所示。途径1：DMDS先在还原酶的作用下被还原为甲硫醇，甲硫醇在氧化酶作用下逐渐被氧化、裂解，最终转化为CO2、SO42-和H2O[21-22]；途径2：DMDS先转化生成二甲硫基甲烷并最终转化为无机产物CO2、SO42-和H2O。
2.5 代谢终产物的测定
利用离子色谱检测菌株R1降解DMDS生成硫酸根的累积量，结果如图10所示。初始DMDS的浓度为16.66 mg/L，待DMDS完全降解后，理论上所有的硫元素都应该转化为SO42-[23]。DMDS的含硫量为53.3%，因此16.66 mg/L的DMDS应转化生成8.704 mg/L的硫酸根。离子色谱检测到硫酸根离子的浓度为5.86 mg/L，因此DMDS完全降解转化生成硫酸根的转化率为65.99%。随着DMDS的降解，SO42-浓度不断增加，但SO42-的产生与DMDS降解之间存在一段延滞期，这是因为DMDS首先被降解为中间产物。在12 h内可检测到甲硫醇，随后才被彻底降解为无机物[24]。
3 讨论
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目前，利用微生物体系降解VOCs污染物已有相关报道[25]，但关于微生物自身分泌表面活性剂降解DMDS的研究较少。本研究从东寨港红树林底泥中分离到一株可以高效降解DMDS且兼具自产表面活性剂功能的菌株R1，并在GenBank中将菌株R1的16S rRNA基因测序列与已知菌种序列比对并构建系统发育树，发现菌株R1与木糖氧化无色杆菌(Achromobacter sp.)具有最高相似性(100%)，表明菌株R1为木糖氧化无色杆菌。在体系温度30 ℃和pH 7.0的条件下，经过120 h降解后，12.49 mg/LDMDS的降解率达到70.74%。Liang等[13]利用蜡样芽孢杆菌(Bacillus cereus) GIGAN2开展生物降解10 mg/L气态DMDS的研究，96 h后去除率可达100%，但生成了二甲基硫和二甲基三硫等中间产物。丁军等[16]利用DMDS降解菌芽孢杆菌属(Bacillus Cohn) SZT-1，在DMDS初始浓度为250 mg/L、温度为30 ℃、pH 5.0的条件下，通过添加淀粉和蛋白胨作为额外的碳源和氮源辅助DMDS的生物利用，经过86.6 h对DMDS的降解率达到50%，但并未对生物降解DMDS的机制进行阐明。
目前已有研究报道，添加表面活性剂可提高生物处理疏水性VOCs的生物利用度。表面活性剂因其具有提高溶解度、降低表面张力、润湿能力及发泡能力等特性而被广泛应用。Li等[26]报道称，在生物滴滤器降解八甲基环四硅氧烷(D4)的过程中，发现铜绿假单胞菌(Pseudomonas aeruginosa) S240产生的生物表面活性剂鼠李糖脂是改善去除D4的主要因素。Tu等[27]报道，生物表面活性剂皂苷可有效增强正己烷的去除效率，并降低生物质在生物滴滤池中的生物量积累速率，表明使用生物表面活性剂能够提高生物处理疏水性VOCs的去除效率。然而，DMDS是一种疏水性VOSCs，其溶解度较低，对细菌种类的毒性较高，且对人体危害较大。本研究表明，微生物菌株R1不仅可以高效降解DMDS，还兼具自产生物表面活性剂的功能。通过乳化能力测定、1H NMR和FT-IR分析，结果表明菌株R1利用DMDS自产的生物表面活性剂与鼠李糖脂的表征结果高度相似，证明该菌株产生的生物表面活性剂为糖脂类。这一结果与Moura等[28]利用铜绿假单胞菌(P. aeruginosa) LBI 2A1分泌的生物表面活性剂种类相似。
Zheng等[29]研究表明甲硫醇是DMDS降解的主要中间产物。本研究通过GC/MS验证，菌株R1降解DMDS的中间产物为二甲硫基甲烷和甲硫醇，推测可能存在2条降解途径，待甲硫醇持续矿化后最终生成硫酸根沉淀及其他无机物，硫元素的转化率为65.99%。硫元素的分析尚未达到平衡，但大部分硫元素已转化为SO42-。Zhao等[30]研究发现，在微生物燃料电池降解DMDS时，DMDS中68.5%的硫元素转化为SO42-，0.08%转化为S2- (或HS-、H2S)，0.7%转化为S0。这些结果表明菌株R1在降解DMDS方面具有较大潜力。
4 结论
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从东寨港红树林底泥中分离到一株可以高效降解DMDS且兼具自产表面活性剂的菌株R1，经16S rRNA基因鉴定该菌为木糖氧化无色杆菌。在降解时间为120 h、温度为30 ℃、DMDS浓度为12.49 mg/L、湿基固体接种量为1.0 g/L的条件下，菌株对DMDS的总降解率为70.74%。经乳化能力测定，1H NMR和FT-IR分析，该菌株产生的生物表面活性剂类型为糖脂类。在不同DMDS浓度范围内，该菌株表现出较好的降解能力和表面活性剂产生性能。经GC/MS验证，主要降解中间产物为甲硫醇和二甲硫基甲烷，S元素转化率为65.99%。
基金
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  • 海南省自然科学基金(425RC684)
  • 国家自然科学基金(32060291)
文献
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2025年第65卷第8期
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doi: 10.13343/j.cnki.wsxb.20250049
  • 接收时间：2025-01-17
  • 首发时间：2026-02-06
  • 出版时间：2025-08-04
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  • 收稿日期：2025-01-17
  • 录用日期：2025-03-05
基金
Hainan Provincial Natural Science Foundation(425RC684)
海南省自然科学基金(425RC684)
National Natural Science Foundation of China(32060291)
国家自然科学基金(32060291)
作者信息
    1.海南大学 生态学院，海南 海口
    2.海南大学 化学化工学院，海南 海口

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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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