Article(id=1241377722417598539, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1241377719049572379, articleNumber=null, orderNo=null, doi=10.13343/j.cnki.wsxb.20230616, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1696694400000, receivedDateStr=2023-10-08, revisedDate=null, revisedDateStr=null, acceptedDate=1708963200000, acceptedDateStr=2024-02-27, onlineDate=1773897112706, onlineDateStr=2026-03-19, pubDate=1717430400000, pubDateStr=2024-06-04, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1773897112706, onlineIssueDateStr=2026-03-19, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1773897112706, creator=13701087609, updateTime=1773897112706, updator=13701087609, issue=Issue{id=1241377719049572379, tenantId=1146029695717560320, journalId=1192105938417971205, year='2024', volume='64', issue='6', pageStart='1691', pageEnd='2143', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1773897111904, creator=13701087609, updateTime=1773897665313, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1241380040286458828, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1241377719049572379, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1241380040286458829, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1241377719049572379, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=2104, endPage=2114, ext={EN=ArticleExt(id=1241377722841223251, articleId=1241377722417598539, tenantId=1146029695717560320, journalId=1192105938417971205, language=EN, title=Identification of aRhodotorula taiwanensis strain capable of removing Mn(Ⅱ), columnId=1241377722715394129, journalTitle=Acta Microbiologica Sinica, columnName=Geomicrobiological Applications, runingTitle=null, highlight=null, articleAbstract=

[Objective] To investigate the removal efficiency and mechanism by a strain ofRhodotorula. [Methods] The strain was identified based on the morphological characteristics and the phylogenetic tree based on 26S rRNA gene sequences. Then, the removal effect of the strain on Mn(Ⅱ) was studied at different initial pH and Mn(Ⅱ) concentrations. Finally, the products were characterized by scanning electron microscopy, X-ray diffraction analysis, and X-ray photoelectron spectroscopy. [Results] The isolate was identified asRhodotorula taiwanensis, which could grow well at pH 2.0 and 2 000 mg/L Mn(Ⅱ). The removal rate of Mn(Ⅱ) by the strain reached 98.52% at the initial pH 6.0 and Mn(Ⅱ) concentration of 300 mg/L after 144 h, while higher concentrations (≥500 mg/L) of Mn(Ⅱ) caused toxicity to the cells and weakened the removal effect. The strain could oxidize Mn(Ⅱ) to manganese oxides (mainly amorphous MnO2, Mn2O3, and MnO), which produced layered minerals accumulated on the cell surface. In addition, the process of removing Mn(Ⅱ) by the strain increased the pH of the medium (initial pH 7.0, Mn(Ⅱ) concentration of 100 mg/L) up to 8.4 after 144 h. [Conclusion] R.taiwanensis MF4 isolated in this study can tolerate low pH and high concentrations of Mn(Ⅱ), remove Mn(Ⅱ), and increase medium pH. The findings have a reference value for the end process design in the remediation and treatment of acid mine drainage.

, correspAuthors=Jin WANG, authorNote=null, correspAuthorsNote=
*WANG Jin, E-mail:
, copyrightStatement=Copyright ©2024 Acta Microbiologica Sinica. All rights reserved., 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=Xiao HE, Zhengbo YUE, Jiashun YIN, Shaoping WANG, Wensheng LIU, Xian'ang ZHANG, Xin CHUAI, Jin WANG), CN=ArticleExt(id=1241377724611219624, articleId=1241377722417598539, tenantId=1146029695717560320, journalId=1192105938417971205, language=CN, title=一株台湾红酵母(Rhodotorula taiwanensis)的鉴定及其除Mn(Ⅱ)性能研究, columnId=1241377722941886549, journalTitle=微生物学报, columnName=地质微生物应用, runingTitle=null, highlight=null, articleAbstract=

【目的】探究一株红酵母对Mn(Ⅱ)的去除效率及其作用机制。【方法】从酸性矿山废水中分离出一株耐酸酵母菌,通过形态和26S rRNA基因测序对菌种进行鉴定,研究不同pH和Mn(Ⅱ)浓度对该菌除Mn(Ⅱ)效果的影响。采用扫描电镜、X射线衍射分析和X射线光电子能谱仪进行产物表征。【结果】分离得到的酵母菌经鉴定为台湾红酵母(Rhodotorula taiwanensis),其在pH 2.0、2 000 mg/L Mn(Ⅱ)条件下仍能生长较好。在初始pH 6.0、Mn(Ⅱ) 300 mg/L条件下培养144 h后, 对Mn(Ⅱ)的去除率能达到98.52%;然而较高浓度的Mn(Ⅱ) (≥500 mg/L)会对细胞产生毒性,从而降低去除效果。R.taiwanensis MF4在去除Mn(Ⅱ)的过程中可以将Mn(Ⅱ)氧化成锰氧化物(主要为无定型的MnO2、Mn2O3、MnO),形成层状物质在细胞表面积累,而且能产生碱度,提升环境pH值,最高可达8.4 [初始pH 7.0,Mn(Ⅱ) 100 mg/L,144 h]。【结论】R.taiwanensis MF4具有耐受低pH和高浓度Mn(Ⅱ)、有效去除Mn(Ⅱ)以及产碱的作用,研究结果对酸性矿山废水修复与治理的末端工艺设计具有参考价值。

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一株台湾红酵母(Rhodotorula taiwanensis)的鉴定及其除Mn(Ⅱ)性能研究
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贺笑 1, 2, 3 , 岳正波 1 , 尹家顺 1 , 王绍平 2 , 刘文胜 3 , 张先昂 4 , 揣新 2 , 王进 1, *
微生物学报 | 地质微生物应用 2024,64(6): 2104-2114
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微生物学报 | 地质微生物应用 2024, 64(6): 2104-2114
一株台湾红酵母(Rhodotorula taiwanensis)的鉴定及其除Mn(Ⅱ)性能研究
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贺笑1, 2, 3, 岳正波1, 尹家顺1, 王绍平2, 刘文胜3, 张先昂4, 揣新2, 王进1, *
作者信息
  • 1 合肥工业大学资源与环境工程学院, 安徽 合肥 230009
  • 2 安徽马钢矿业资源集团南山矿业有限公司, 安徽 马鞍山 243000
  • 3 安徽马钢矿业资源集团有限公司, 安徽 马鞍山 243000
  • 4 安徽马钢矿业资源集团生态修复科技分公司, 安徽 马鞍山 243000
Identification of aRhodotorula taiwanensis strain capable of removing Mn(Ⅱ)
Xiao HE1, 2, 3, Zhengbo YUE1, Jiashun YIN1, Shaoping WANG2, Wensheng LIU3, Xian'ang ZHANG4, Xin CHUAI2, Jin WANG1, *
Affiliations
  • 1 School of Resources and Environmental Engineering, Hefei University of Technology, Hefei 230009, Anhui, China
  • 2 Anhui Masteel Mining Resources Group, Nanshan Mining Co., Ltd., Ma'anshan 243000, Anhui, China
  • 3 Anhui Masteel Mining Resources Group Co., Ltd., Ma'anshan 243000, Anhui, China
  • 4 Ecological Restoration Technology Branch of Anhui Masteel Mining Resources Group Co., Ltd., Ma'anshan 243000, Anhui, China
出版时间: 2024-06-04 doi: 10.13343/j.cnki.wsxb.20230616
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【目的】探究一株红酵母对Mn(Ⅱ)的去除效率及其作用机制。【方法】从酸性矿山废水中分离出一株耐酸酵母菌,通过形态和26S rRNA基因测序对菌种进行鉴定,研究不同pH和Mn(Ⅱ)浓度对该菌除Mn(Ⅱ)效果的影响。采用扫描电镜、X射线衍射分析和X射线光电子能谱仪进行产物表征。【结果】分离得到的酵母菌经鉴定为台湾红酵母(Rhodotorula taiwanensis),其在pH 2.0、2 000 mg/L Mn(Ⅱ)条件下仍能生长较好。在初始pH 6.0、Mn(Ⅱ) 300 mg/L条件下培养144 h后, 对Mn(Ⅱ)的去除率能达到98.52%;然而较高浓度的Mn(Ⅱ) (≥500 mg/L)会对细胞产生毒性,从而降低去除效果。R.taiwanensis MF4在去除Mn(Ⅱ)的过程中可以将Mn(Ⅱ)氧化成锰氧化物(主要为无定型的MnO2、Mn2O3、MnO),形成层状物质在细胞表面积累,而且能产生碱度,提升环境pH值,最高可达8.4 [初始pH 7.0,Mn(Ⅱ) 100 mg/L,144 h]。【结论】R.taiwanensis MF4具有耐受低pH和高浓度Mn(Ⅱ)、有效去除Mn(Ⅱ)以及产碱的作用,研究结果对酸性矿山废水修复与治理的末端工艺设计具有参考价值。

锰离子  /  红酵母  /  除锰机制  /  产碱  /  酸性矿山废水

[Objective] To investigate the removal efficiency and mechanism by a strain ofRhodotorula. [Methods] The strain was identified based on the morphological characteristics and the phylogenetic tree based on 26S rRNA gene sequences. Then, the removal effect of the strain on Mn(Ⅱ) was studied at different initial pH and Mn(Ⅱ) concentrations. Finally, the products were characterized by scanning electron microscopy, X-ray diffraction analysis, and X-ray photoelectron spectroscopy. [Results] The isolate was identified asRhodotorula taiwanensis, which could grow well at pH 2.0 and 2 000 mg/L Mn(Ⅱ). The removal rate of Mn(Ⅱ) by the strain reached 98.52% at the initial pH 6.0 and Mn(Ⅱ) concentration of 300 mg/L after 144 h, while higher concentrations (≥500 mg/L) of Mn(Ⅱ) caused toxicity to the cells and weakened the removal effect. The strain could oxidize Mn(Ⅱ) to manganese oxides (mainly amorphous MnO2, Mn2O3, and MnO), which produced layered minerals accumulated on the cell surface. In addition, the process of removing Mn(Ⅱ) by the strain increased the pH of the medium (initial pH 7.0, Mn(Ⅱ) concentration of 100 mg/L) up to 8.4 after 144 h. [Conclusion] R.taiwanensis MF4 isolated in this study can tolerate low pH and high concentrations of Mn(Ⅱ), remove Mn(Ⅱ), and increase medium pH. The findings have a reference value for the end process design in the remediation and treatment of acid mine drainage.

manganese ion  /  Rhodotorula taiwanensis  /  manganese removal mechanism  /  alkali production  /  acid mine drainage
贺笑, 岳正波, 尹家顺, 王绍平, 刘文胜, 张先昂, 揣新, 王进. 一株台湾红酵母(Rhodotorula taiwanensis)的鉴定及其除Mn(Ⅱ)性能研究. 微生物学报, 2024 , 64 (6) : 2104 -2114 . DOI: 10.13343/j.cnki.wsxb.20230616
Xiao HE, Zhengbo YUE, Jiashun YIN, Shaoping WANG, Wensheng LIU, Xian'ang ZHANG, Xin CHUAI, Jin WANG. Identification of aRhodotorula taiwanensis strain capable of removing Mn(Ⅱ)[J]. Acta Microbiologica Sinica, 2024 , 64 (6) : 2104 -2114 . DOI: 10.13343/j.cnki.wsxb.20230616
在硫化矿物开采过程中,尾矿和弃土被堆积在露天场地,长期暴露于自然环境中,这些废弃物在空气、水和微生物的共同作用下形成了酸性矿山废水(acid mine drainage, AMD)。AMD具有低pH值,并含有大量金属离子,其中Mn(Ⅱ)浓度一般在20−200 mg/L[1],有些能达到500 mg/L[2-3]甚至是1 000 mg/L以上[4],再加上其特殊的化学性质使其治理难度较大[5]。首先,锰的化学中和处理方法需要较高的pH环境(pH > 9.0),需要大量的碱性物质,导致处理成本高且易产生二次废物[6];其次,水溶液中的其他金属离子会抑制Mn(Ⅱ)的氧化速率[7];再者,由于Mn(Ⅱ)的硫化物相具有较大的溶解度,使其相对于其他过渡金属更难去除[8]
在自然环境中,微生物可以通过直接途径(酶促反应等)和间接途径(改变pH值、氧化还原电位等)对Mn(Ⅱ)进行氧化,该过程是控制环境中Mn氧化物形成和生物地球化学循环的主导因素[9],其氧化反应速率比非生物过程高数个数量级,因此,微生物方法为废水中Mn(Ⅱ)的去除提供了新的选择,如利用细菌[10]、真菌[11]和藻类[1]等微生物实现生物脱锰。其中,酵母菌因其经济、安全和对金属离子吸附能力强等优势,具有较高的潜在应用价值[12]
近年来,对酵母菌在Mn(Ⅱ)吸附及去除机制方面的研究逐渐深入,涉及多个酵母菌属种,不同种类的酵母对Mn(Ⅱ)的去除机制各异。研究发现,酵母细胞可以通过吸附、积累、氧化等方式对环境中的Mn(Ⅱ)进行去除[13]。酵母细胞壁中的甘露聚糖以及细胞表面蛋白质中游离的羧基、羟基、胺、磷酸盐和氢硫化物基团等均可结合金属离子,因此酵母菌是一种高效的生物吸附剂[14-15]。当金属离子穿过屏障进入细胞后,会在细胞中积累,一些酵母菌还会释放酶类物质(如多铜氧化酶、MCOs)将Mn(Ⅱ)氧化成MnOx[16]。已有报道发现,从植物中分离出的一种野生酵母Rhodotorula taiwanensis,能够在低pH值(2.3)、高浓度汞铬化合物或Al3+ (200 mmol/L)存在的条件下生长,在极端酸性环境条件中可形成生物膜[17]。然而,关于R.taiwanensis对酸性废水中Mn(Ⅱ)的去除及其作用机制,目前仍缺乏深入的研究。这些研究结果表明该种在酸性、金属含量高的废水修复中具有良好的应用前景。
微生物对金属离子的去除性能与其接触的微环境密切相关,包括溶液的初始pH值和金属离子浓度等因素。多数真菌对重金属富集的最适pH值为5.0或6.0;当pH值大于5.0时,酿酒酵母的生物吸附效果最佳[18]。除此之外,溶液中重金属的浓度会直接影响微生物的生长代谢及其对重金属的吸附效率[19]。因此,本研究以从安徽某矿山酸性废水坑湖中筛选出的R.taiwanensis为研究对象,考察环境pH和初始Mn(Ⅱ)浓度对其除锰效率的影响,结合产物形貌、物质形态与化学组成,揭示该菌株的除锰特性,从而为深入认识该菌株在酸性矿山废水中的生物修复作用提供研究基础和科学依据。
水样采自安徽省东部某矿山酸性坑湖,该坑湖pH值较低(3.0−3.8),锰浓度较高,最高可达800 mg/L[20]。取其水样50 mL,3 000 r/min离心5 min后,取1 mL上清液接种至50 mL酵母浸出粉胨葡萄糖培养基(yeast extract peptone dextrose, YPD)中,于28 ℃、120 r/min富集培养72 h。对富集后的样品在孟加拉红固体培养基(RBA) [含Mn(Ⅱ) 300 mg/L,pH 5.0]上进行分离纯化,反复划线筛选,获得一株菌株记为菌株MF4。
分离纯化后的菌株MF4在高倍显微镜下进行观察,记录真菌颜色、大小和形状。
利用Ezup柱式真菌基因组DNA抽提试剂盒[生工生物工程(上海)股份有限公司]提取MF4全基因组DNA。利用通用引物对NL1 (5′-GCATATCAATAAGCGGAGGAAAAG-3′)和NL4 (5′-GGTCCGTGTTTCAAGACGG-3′)扩增其26S rRNA基因序列片段(25 μL体系)。PCR扩增体系和程序同Miao等[21]。扩增后采用凝胶电泳法检测扩增结果并送至生工生物工程(上海)股份有限公司进行测序。
测序结果提交至NCBI (https://blast.ncbi.nlm.nih.gov/Blast.cgi)进行比对分析,下载亲缘关系相近种群的26S rRNA基因序列片段,在MEGA (version 6)中比对校正后,采用邻接法(neighbor-joining, NJ)构建系统发育树,确定菌株MF4基于分子生物学的分类地位。
为了研究酵母菌的耐酸、耐锰特性,分别将0.082 g/L生物量的酵母菌接种到不同pH (2.0、3.0、4.0、5.0、6.0和7.0)和不同Mn(Ⅱ)浓度(50、100、300、500、800、1 000和2 000 mg/L)的培养基中,于30 ℃、120 r/min培养144 h后取样称量酵母菌干重。
为了评估初始pH对酵母菌除锰效率的影响,将0.082 g/L生物量的酵母接种到含有50 mg/L[15] Mn(Ⅱ) (MnSO4)培养基中,初始pH设置为4.0、5.0、5.5、6.0、7.0,每个实验条件设置不加菌空白对照,于30 ℃、120 r/min培养144 h,每24 h取样测定培养基pH值,原子吸收光谱法分析上清液中Mn(Ⅱ)浓度。
在研究初始Mn(Ⅱ)浓度对酵母菌除锰效率影响的研究中,选择在50 mg/L Mn(Ⅱ)溶液中去除效果较好的pH条件,根据酵母菌的耐受特性和研究应用场景,Mn(Ⅱ)初始浓度分别设置为50、100、300、500、800 mg/L,每个实验条件设置不加菌空白对照,于30 ℃、120 r/min培养144 h后取样,测定培养基pH值,原子吸收光谱法分析上清液中Mn(Ⅱ)浓度。
将培养完成后的菌液离心、洗涤后冷冻干燥,并研磨获得固体粉末。固体产物采用扫描电镜(scanning electron microscope, SEM)分析细胞形貌特征;采用X射线光电子能谱仪(X-ray photoelectron spectrometer, XPS) (ThermoFisher Scientific)分析锰价态组成;X射线衍射(X-ray diffraction, XRD)分析的样品在2θ范围从5°到70°范围内扫描。
菌株MF4的形态多呈球形、卵圆形,一般为1−5 μm或5−20 μm,无鞭毛,不能游动(图1A),菌落大而厚,菌落表面光滑、湿润、黏稠,容易挑起,菌落质地均匀,红色,正反面、边缘及中央部位的颜色均一(图1B)。
在实验过程中,随着培养时间的延长,对照组[未添加Mn(Ⅱ)]培养基为橙红色(图1C),而实验组[添加Mn(Ⅱ)]培养基的颜色呈现棕色(图1D)。
26S rRNA基因序列测序比对结果如图2所示,菌株MF4隶属于红酵母属Rhodotorula,与R.taiwanensis种菌株(相似性为100%)聚为一支,MF4应初步鉴定为R.taiwanensis,命名为Rhodotorula taiwanensis MF4。
不同pH条件下R.taiwanensis MF4的生长情况如图3A所示,在较宽的pH范围内(pH 2.0−7.0)R.taiwanensis MF4均可生长,pH 2.0条件下生物量虽然显著低于pH 3.0−7.0条件,但是仍然生长较好,7 d后的生物量能达到(7.38±0.24) g/L。pH≥3.0条件下,各组之间无显著差异,生物量能达到(8.70±0.20) g/L。不同Mn(Ⅱ)浓度条件下R.taiwanensis MF4的生长情况如图3B所示,培养7 d后,红酵母均能较好地生长,当Mn(Ⅱ)浓度为100 mg/L时生长最好,7 d后的生物量为(10.91±0.47) g/L;当Mn(Ⅱ)浓度为300−800 mg/L时,生物量为(10.11±0.27) g/L,与50 mg/L时(10.25±0.72) g/L的生物量之间无显著差异。当Mn(Ⅱ)浓度达到1 000 mg/L以后,生物量显著降低[(9.31±0.38) g/L,P < 0.01],Mn(Ⅱ)浓度达到2 000 mg/L时的生物量仍然能达到较好条件下的80% [(8.90±0.03) g/L]。结果表明红酵母是一株耐酸、耐Mn(Ⅱ)真菌,pH≥3.0的条件更适于该酵母生长,即使Mn(Ⅱ)浓度达到2 000 mg/L也能良好生长。
初始pH对R.taiwanensis MF4除锰效率的影响研究结果如图4所示,在初始50 mg/L Mn(Ⅱ)条件下,初始pH值较低时(4.0或5.0),R.taiwanensis MF4对Mn(Ⅱ)的去除作用不明显。当初始pH≥5.5时,培养48 h后,培养基的pH呈现上升趋势,同时Mn(Ⅱ)的去除效率也在增长。当培养基pH值增长到6.6时(96 h,初始pH 7.0条件下),Mn(Ⅱ)的去除效率显著提高至(64.16±7.66)%,而在初始pH值为5.5和6.0的条件下,此时的培养基pH值分别为6.0和6.2,Mn(Ⅱ)的去除效率并没有显著提高[初始pH 5.5的去除率为(18.16±2.54)%,初始pH 6.0的去除率为(16.80±2.93)%]。培养120 h后,培养基pH值分别达到7.2、7.5和7.4 (初始pH分别为5.5、6.0和7.0),Mn(Ⅱ)的去除效率显著提高到(89.46±1.42)%、(88.97±0.40)%和(90.91±1.36)%,培养后期,锰的最大去除率能达到93.61% (初始pH 7.0)。空白无菌对照组的pH在不同时期均无明显变化,对照组中未观察到Mn(Ⅱ)去除。
本研究中,初始pH较低的条件下(4.0或5.0),去除效果不明显的原因可能为:首先,大量氢离子的存在,使细胞表面带正电荷,导致官能团与Mn(Ⅱ)产生静电排斥作用[22];其次,氢离子与Mn(Ⅱ)同时竞争与细胞表面官能团结合的机会,使细胞对Mn(Ⅱ)的吸附作用较弱;再者,已有研究表明酵母菌对Mn(Ⅱ)的去除作用与自身的代谢有关[23],在环境pH较低的情况下,细胞代谢功能较差,从而使生物去除作用较弱[24],当环境pH能够保持在7.0−7.5时,锰的生物氧化作用更容易进行[25]。本研究中,培养120 h后,培养基pH分别达到7.2、7.5和7.4,去除效率显著提高(图4)。
另一方面,即使在合适pH条件下(≥5.5),R.taiwanensis MF4对Mn(Ⅱ)的去除率在培养96 h后(初始pH 7.0)才开始显著提高,表明R.taiwanensis MF4的吸附活性和氧化活性在生长过程中可能发挥着不同的作用[26],接触时间的延长有利于Mn(Ⅱ)的去除。在生长初期,R.taiwanensis MF4未对环境作出反应,氧化活性低,只发生了少量的吸附作用,导致Mn(Ⅱ)的去除率较低,当酵母细胞适应环境后,不断输送酶类物质参与Mn(Ⅱ)氧化,从而提高了Mn(Ⅱ)的去除效率[27]。本研究中,随着培养时间的延长,Mn(Ⅱ)去除率相比于红酵母属另一菌株R.mucilaginosa在同等条件下对Mn(Ⅱ)去除效率高了近9倍[12]。以上结果表明R.taiwanensis MF4在初始50 mg/L Mn(Ⅱ)、初始pH≥5.5条件下,对Mn(Ⅱ)具有较好的去除效果。
培养144 h后,在去除效果较好的初始pH (pH≥5.5)和不同初始Mn(Ⅱ)浓度培养条件下,红酵母对Mn(Ⅱ)的去除效率如图5所示。总体而言,该菌株对Mn(Ⅱ)具有较好的耐受性和去除效率。当Mn(Ⅱ)浓度≤300 mg/L时,该菌株在不同初始pH (5.5、6.0、7.0)条件下对Mn(Ⅱ)的去除率无显著差异,均可超过90.00%,最大值为98.52% [初始Mn(Ⅱ) 300 mg/L,初始pH 6.0]。当初始Mn(Ⅱ)浓度为500 mg/L、初始pH 7.0条件下,R.taiwanensis MF4对Mn(Ⅱ)的去除效率[(62.79±0.91)%]显著高于酸性条件下[(8.73±4.74)%,P < 0.05)。当Mn(Ⅱ)为800 mg/L时,该菌株在中性和酸性条件下的去除效果均较差(< 10%)。
图5所示,初始Mn(Ⅱ)浓度≤300 mg/L时,各组对Mn(Ⅱ)的去除效果无显著差异。当Mn(Ⅱ)初始浓度从300 mg/L分别升高至500 mg/L和800 mg/L时,R.taiwanensis MF4对Mn(Ⅱ)的去除率从(94.72±2.55)%分别降低至(26.75± 28.15)% (P < 0.05)和(5.42±3.41)% (P < 0.05),这与芽孢杆菌(Bacillus cereus) HM-5对锰的去除行为类似[28]。结果表明,R.taiwanensis MF4对低浓度Mn(Ⅱ) (< 300 mg/L)具有较好的耐受性和较高的去除率,而高浓度的Mn(Ⅱ)会对细胞产生毒性,进而抑制了对Mn(Ⅱ)的去除作用。
本研究培养基终点的pH如图6所示,除初始Mn(Ⅱ)浓度为800 mg/L组,其余各组培养终点的pH值均高于初始值。当初始Mn(Ⅱ)浓度为50−300 mg/L时,初始pH值对培养终点的pH值无显著影响(均值7.8±0.3),最高可达到8.4 [初始pH 7.0,Mn(Ⅱ) 100 mg/L],最高增长率为52% [初始pH 5.5,Mn(Ⅱ) 100 mg/L,终点pH 8.4],显著高于初始Mn(Ⅱ)浓度为500 mg/L (均值6.7±1.3)和800 mg/L (均值4.7±0.6)培养基的pH值。结果表明,R.taiwanensis MF4在积累Mn(Ⅱ)的过程中会产生碱度,提升环境pH值,而较高的Mn(Ⅱ)浓度会影响酵母菌的代谢活动从而使产碱能力下降。Amorim等[23]在研究Meyerozyma属2个菌株对锰的耐受性和吸附实验中观察到了酵母菌产碱的现象。酵母在生长前期,会产生有机酸,使培养基pH值降低(图4),发酵后期,pH值的升高可能与培养基中有机物的组成[碳氮比(C/N)]有关[29]
图5所示,初始Mn(Ⅱ) < 300 mg/L时,酵母菌R.taiwanensis MF4对锰具有较好的去除效果,且培养后期含Mn(Ⅱ)培养基的颜色变成棕色(图1D),表明R.taiwanensis MF4在去除Mn(Ⅱ)的过程中可能产生了生物氧化作用,形成了生物锰氧化物[30]。利用扫描电镜(SEM)观察300 mg/L Mn(Ⅱ)试验组的固相产物,如图7A所示,细胞表面形成层状物质,类似于在对其他细菌和真菌的生物氧化固相产物中观察到的层状锰氧化物[31]
进一步通过XRD衍射分析初步考察产物表面物质组成。真菌锰氧化物通常包含层状结构的锰酸盐δ-MnO2、α-MnOOH和一些固体伴生锰(Ⅱ)[8]。不同种类的Mn(Ⅱ)氧化真菌会产生结构多样性的矿物质。由于真菌源性锰氧化物晶型较差,在图谱中的衍射峰均不显著(图7B),在2θ处,22.4°、34.5°、37.0°、38.4°、40.8°、42.4°、56.0°、57.6°和65.7°的衍射峰与MnO2 (JCPDS: 12-0713)的晶相相对应。在2θ为23.2°、26.8°、33.0°、35.7°、38.2°、40.7°、43.0°、49.5°、53.5°、64.2°和65.7°处的峰分别对应于Mn2O3 (JCPDS: 02-0902)的(211)、(220)、(222)、(321)、(400)、(411)、(420)、(431)、(521)、(541)和(622)晶格面。在2θ为34.9°、40.5°、58.7°处的衍射峰对应于MnO (JCPDS: 06-0592)的(111)、(200)、(220)晶格面。因此XRD图谱比对出氧化物中存在的物质主要有MnO2、Mn2O3和少量的MnO。
XPS光谱分析结果表明(图7C7D),C、N、O和Mn是酵母细胞表面的主要元素,大量O和Mn的存在表明细胞表面有MnOx的沉积[12],在Mn 2p高分辨率光谱中,在642 eV处的峰归属于Mn 2p3/2,氧化产物为混价态锰化合物;在640.81 eV的峰归属于Mn(Ⅱ),其占比为14.85%;在641.88 eV的峰归属于Mn(Ⅲ),其占比为54.23%,在643.40 eV的峰归属于Mn(Ⅳ),其占比为30.92%,在647.25 eV处的峰是卫星峰。
以上表征结果表明R.taiwanensis MF4与锰氧化细菌类似[27],能够氧化Mn(Ⅱ)形成锰氧化物沉积在细胞表面。这些锰氧化物具有较大的比表面积,通常与金属离子具有较高的亲和力[32],可以通过共沉淀、吸附、氧化还原过程,在许多重金属污染物的迁移和转化过程中发挥重要作用[33]。然而由于真菌对锰的氧化还原反应是复杂的,涉及多个过程,如通过细胞酶的活性[34]、胞外电子[35]或者非酶蛋白(如boxA)[36]均能参与氧化过程,所以目前仍不知道这种氧化作用是如何产生的,有待深入解析。
本研究分离筛选出一株耐酸红酵母菌株R.taiwanensis MF4,该酵母对Mn(Ⅱ)具有较好去除效果,在初始pH 6.0、Mn(Ⅱ) 300 mg/L条件下,对Mn(Ⅱ)的去除率能达到98.52%。该酵母通过吸附和氧化的方式对溶液中的Mn(Ⅱ)进行去除,将Mn(Ⅱ)氧化成锰氧化物(主要为无定型的MnO2、Mn2O3、MnO),形成层状物质在细胞表面积累,同时产生碱性物质,提高环境pH,最高可达8.4。本研究结果表明R.taiwanensis MF4在去除酸性废水中的Mn(Ⅱ)、提高水环境pH方面具有潜在应用价值,可以为酸性矿山废水的修复和治理工作提供参考。
  • 国家自然科学基金(U20A20325)
  • 安徽省重点研究与开发计划(2022107020015)
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2024年第64卷第6期
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doi: 10.13343/j.cnki.wsxb.20230616
  • 接收时间:2023-10-08
  • 首发时间:2026-03-19
  • 出版时间:2024-06-04
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  • 收稿日期:2023-10-08
  • 录用日期:2024-02-27
基金
National Natural Science Foundation of China(U20A20325)
国家自然科学基金(U20A20325)
Key Research and Development Plan of Anhui Province(2022107020015)
安徽省重点研究与开发计划(2022107020015)
作者信息
    1 合肥工业大学资源与环境工程学院, 安徽 合肥 230009
    2 安徽马钢矿业资源集团南山矿业有限公司, 安徽 马鞍山 243000
    3 安徽马钢矿业资源集团有限公司, 安徽 马鞍山 243000
    4 安徽马钢矿业资源集团生态修复科技分公司, 安徽 马鞍山 243000

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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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