Microbial Mechanism of Electron Shuttle-mediated Antimony Reduction and Mineralization by Soil Microorganism

doi:10.16258/j.cnki.1674-5906.2024.02.011

Ecology and Environment ›› 2024, Vol. 33 ›› Issue (2): 272-281.DOI: 10.16258/j.cnki.1674-5906.2024.02.011

• Research Article [Environmental Sciences] • Previous Articles Next Articles

Microbial Mechanism of Electron Shuttle-mediated Antimony Reduction and Mineralization by Soil Microorganism

LAN Jun¹^,²(), CHEN Guanhong²^,^*(), ZHANG Juntao³, HEMMAT-JOU Mohammad Hossein², SHU Xiaohua¹, FANG Liping², LI Fangbai²

1. College of Environmental Science and Engineering, Guilin University of Science and Technology, Guilin 541000, P. R. China
2. Institute of Eco-environmental and Soil Sciences, Guangdong Academy of Sciences/Guangdong Key Laboratory of Integrated Agro-environmental Pollution Control and Management, Guangzhou 510650, P. R. China
3. Guangzhou Institute of Forestry and Landscape Architecture, Guangzhou 510405, P. R. China

Received:2023-12-03 Online:2024-02-18 Published:2024-04-03

电子穿梭体介导土壤锑还原成矿的微生物机制

蓝浚¹^,²(), 陈冠虹²^,^*(), 张俊涛³, Hemmat-Jou Mohammad Hossein², 舒小华¹, 方利平², 李芳柏²

1.桂林理工大学环境科学与工程学院，广西桂林 541000
2.广东省科学院生态环境与土壤研究所/广东省农业环境综合治理重点实验室，广东广州 510650
3.广州市林业和园林科学研究院，广东广州 510405

通讯作者: *陈冠虹。E-mail: ghchen@soil.gd.cn
作者简介:蓝浚（2000年生），男，硕士研究生，研究方向为土壤锑污染控制。E-mail: lanjun9527@163.com
基金资助:
国家自然科学基金项目(42077354);国家自然科学基金项目(42377395);广州市科技计划项目“广州市生态园林科技协同创新中心”(202206010058);韶关市省科技专项资金项目(220716096270196);韶关市省科技创新战略专项项目(230225176275072)

Abstract

Abstract:

Microbially mediated antimonate [Sb(V)] reduction and subsequent mineralization decrease the availability and mobility of Sb, which is one of the strategies for the remediation of Sb-contaminated soils. Electron shuttle (ES) AQDS has been reported to accelerate Sb reduction by soil microbial enrichment, probably due to stimulated microbial respiration and growth. Understanding the microbial mechanisms underlying Sb reduction is important for controlling Sb contamination in soil. Despite the critical role of ES in altering the electron transport pathway in respiratory metabolism, ES-mediated Sb reduction and microbial transcriptional responses during this process remain unclear. In this study, Mesobacillus jeotgali PS1, a facultative anaerobic Sb-reducing bacterium, was isolated from Sb-contaminated paddy soil to investigate the effects of electron shuttles (AQDS and FMN) on microbial Sb reduction and transcriptional activities of functional genes. The results showed that the dissolved Sb(III) concentration first increased and then decreased over time during the Sb reduction. The efficiency of dissolved Sb removal was 64% after 72 h of incubation and cuboctahedral Sb₂O₃ was produced, suggesting that Sb reduction and mineralization by strain PS1 can lead to Sb passivation. Electron shuttles accelerated Sb reduction and did not affect the crystalline structure or morphology of the mineral products. Quantitative analysis of the transcript abundances of functional genes in strain PS1 influenced by ES revealed that AQDS treatment resulted in a relatively greater upregulation of cytosolic dimethyl sulfoxide reductase (DMSOR) family genes and intracellular detoxifying arsenic reduction gene (arsC) in strain PS1 than that with FM, which could result in greater respiratory metabolic activity and intracellular Sb detoxification, thereby promoting microbial Sb reduction. This study revealed the microbial mechanism of electron shuttle-mediated antimony reduction and mineralization by soil microorganisms, which may be useful for improving bioremediation of Sb-contaminated paddy fields.

Key words: antimony-reducing bacteria, electron shuttle, senarmontite, antimony detoxification, transcriptional responses, dimethyl sulfoxide reductase

摘要：

微生物锑还原成矿有助于降低土壤锑的生物有效性和移动性，是土壤锑污染修复的重要策略之一。电子穿梭体AQDS能够加速土壤富集菌群锑还原速率，可能与其促进微生物呼吸及细胞生长有关。理解电子穿梭体（ES）介导微生物锑还原过程与机制可为土壤锑污染控制提供关键理论支撑。醌类和黄素类电子穿梭体（AQDS和FMN）存在时可能改变微生物呼吸代谢中的电子传递过程，然而ES介导下的微生物锑还原过程及转录响应机制尚不清楚。利用锑污染稻田土壤分离的兼性厌氧锑还原细菌Mesobacillus jeotgali PS1作为研究对象，探究醌类和黄素类电子穿梭体（AQDS和FMN）对菌株PS1锑还原过程及关键功能基因转录活性的影响。结果表明，菌株PS1驱动Sb(V)还原为Sb(III)过程中水溶态Sb(III) 随培养时间先累积后下降，培养72 h后水溶态锑去除率为64%，生成十四面体方锑矿，表明菌株PS1驱动锑还原成矿有助于锑的钝化。两种ES能够加速细菌锑还原反应，而对胞外生成的方锑矿晶型没有影响。通过定量分析菌株PS1潜在功能基因转录表达活性，结果表明AQDS相比FMN更能促进菌株PS1细胞膜二甲基亚砜还原酶（DMSOR）基因和胞内解毒型砷还原基因（arsC）转录活性，有助于增加菌株PS1呼吸代谢活性和胞内锑解毒从而加速锑还原，所以AQDS可能在强化微生物锑还原钝化中更具有优势。该研究揭示了不同电子穿梭体介导锑还原的微生物机制，为锑污染土壤生物修复提供重要理论支撑。

关键词: 锑还原细菌, 电子穿梭体, 方锑矿, 锑解毒, 转录响应, 二甲基亚砜还原酶

CLC Number:

X172

LAN Jun, CHEN Guanhong, ZHANG Juntao, HEMMAT-JOU Mohammad Hossein, SHU Xiaohua, FANG Liping, LI Fangbai. Microbial Mechanism of Electron Shuttle-mediated Antimony Reduction and Mineralization by Soil Microorganism[J]. Ecology and Environment, 2024, 33(2): 272-281.

蓝浚, 陈冠虹, 张俊涛, Hemmat-Jou Mohammad Hossein, 舒小华, 方利平, 李芳柏. 电子穿梭体介导土壤锑还原成矿的微生物机制[J]. 生态环境学报, 2024, 33(2): 272-281.

Figures/Tables 9

Table 1 Details of primer pairs and thermal cycling parameters for qPCR

目的基因	引物名称	引物序列 (5′-3′)	扩增条件
gyrB	gyrB-F	GGCGGTACACACGAATTTGG	95 ℃预变性 2 min; 95 ℃ 10 s, 60 ℃ 20 s, 40个循环
gyrB	gyrB-R	CTTCACGCACGTCTTCTCCT
dmsB	dmsB-F	CCAGATCACGGATGAGGGCG
dmsB	dmsB-R	CGCGTCACTTGGGCAAACAG
nrfC	nrfC-F	GTAAGTGTCTGCCCGACCAA
nrfC	nrfC-R	TACCTTCTGCGACCAACTCG
nasA	nasA-F	CCTGCTACAACATGGGCAGA
nasA	nasA-R	CTGGCCTTTACCGAGACGTT
fdnG	fdnG F	CATCCCACCAAATGACAGCC
fdnG	fdnG-R	GCAAGAACCGCTCCAAGAAC
arsC	arsC-F	AGGCGATGAATGAGGTGGGA
arsC	arsC-R	GTTACCGGGCAGTGCTCATC

Figure 1 Growth curve and SEM image of antimonate-reducing strain PS1 in the medium supplemented with lactate and Sb(V) as electron donor and acceptor

Figure 2 Concentrations of Sb(V), Sb(III) and lactate over time in strain PS1 cultured for 72 h influenced by electron shuttles

Figure 3 XRD patterns of antimony reduction products from strain PS1 after 72 h incubation in Sb, Sb+AQDS and Sb+FMN treatments

Figure 4 SEM-EDS plots of antimony reduction products of strain PS1 after 72 h incubation in Sb, Sb+AQDS and Sb+FMN treatments

Figure 5 TEM-EDS plots of cell-mineral precipitation of strain PS1 after 72 h incubation in Sb, Sb+AQDS and Sb+FMN treatment

Figure 6 Variability in protein amount of strain PS1 after 72 h incubation in CK, Sb, Sb+AQDS and Sb+FMN treatments

Figure 7 Transcriptional responses of DMSOR family genes and arsenic resistance genes in strain PS1 under different treatments (Fc is the variance multiplier)

Figure 8 Schematic diagram of electron shuttles mediating antimony reduction process in strain PS1

References 40

[1]	ABIN C A, HOLLIBAUGH J T, 2013. Dissimilatory antimonate reduction and production of antimony trioxide microcrystals by a novel microorganism[J]. Environmental Science & Technology, 48(1): 681-688. DOI URL
[2]	ABIN C A, HOLLIBAUGH J T, 2019. Transcriptional response of the obligate anaerobe Desulfuribacillus stibiiarsenatis MLFW-2T to growth on antimonate and other terminal electron acceptors[J]. Environmental Microbiology, 21(2): 618-630. DOI URL
[3]	BOLAN N, KUMAR M, SINGH E, et al., 2022. Antimony contamination and its risk management in complex environmental settings: A review[J]. Environment International, 158: 106908. DOI URL
[4]	BUTCHER BRONWYN G, DEANE SHELLY M, RAWLINGS DOUGLAS E, 2000. The chromosomal arsenic resistance genes of thiobacillus ferrooxidans have an unusual arrangement and confer increased arsenic and antimony resistance to Escherichia coli[J]. Applied and Environmental Microbiology, 66(5): 1826-1833. DOI URL
[5]	FILELLA M, BELZILE N, CHEN Y W, 2002. Antimony in the environment: a review focused on natural waters: II. Relevant solution chemistry[J]. Earth-Science Reviews, 59(1): 265-285. DOI URL
[6]	GUO X J, WU Z J, HE M C, et al., 2014. Adsorption of antimony onto iron oxyhydroxides: Adsorption behavior and surface structure[J]. Journal of Hazardous Materials, 276: 339-345. DOI PMID
[7]	HARSHITHA R, ARUNRAJ D R, 2021. Real-time quantitative PCR: A tool for absolute and relative quantification[J]. Biochemistry and Molecular Biology Education, 49(5): 800-812. DOI URL
[8]	HE M C, YANG J R, 1999. Effects of different forms of antimony on rice during the period of germination and growth and antimony concentration in rice tissue[J]. Science of the Total Environment, 243-244: 149-155. DOI URL
[9]	JORMAKKA M, TöRNROTH S, BYRNE B, et al., 2002. Molecular basis of proton motive force generation: Structure of formate dehydrogenase-N[J]. Science, 295(5561): 1863-1868. PMID
[10]	KIELKOPF C L, BAUER W, URBATSCH I L, 2020. Bradford assay for determining protein concentration[M]. Cold Spring Harbor Protocols, (4): 102269.
[11]	KLÜPFEL L, PIEPENBROCK A, KAPPLER A, et al., 2014. Humic substances as fully regenerable electron acceptors in recurrently anoxic environments[J]. Nature Geoscience, 7(3): 195-200. DOI
[12]	LI Q, HUANG M H, SHU S H, et al., 2022. Quinone-mediated Sb removal from sulfate-rich wastewater by anaerobic granular sludge: Performance and mechanisms[J]. Science of The Total Environment, 838(Part 3): 156217. DOI URL
[13]	MENG Y L, LIU Z J, ROSEN B P, 2004. As(III) and Sb(III) uptake by GlpF and efflux by ArsB in Escherichia coli[J]. Journal of Biological Chemistry, 279(18): 18334-18341. DOI URL
[14]	MORENO-VIVIáN C, FLORES E, 2007. Chapter 17 - nitrate assimilation in bacteria[M]// Biology of the Nitrogen Cycle. Amsterdam: Elsevier: 263-282.
[15]	MURCIEGO A M, SÁNCHEZ A G, GONZÁLEZ M A R, et al., 2007. Antimony distribution and mobility in topsoils and plants (Cytisus striatus, Cistus ladanifer and Dittrichia viscosa) from polluted Sb-mining areas in Extremadura (Spain)[J]. Environmental Pollution, 145(1): 15-21. DOI PMID
[16]	NGUYEN V K, PARK Y, LEE T, 2019. Microbial antimonate reduction with a solid-state electrode as the sole electron donor: A novel approach for antimony bioremediation[J]. Journal of Hazardous Materials, 377: 179-185. DOI PMID
[17]	OKAMOTO A, SAITO K, INOUE K, et al., 2014. Uptake of self-secreted flavins as bound cofactors for extracellular electron transfer in Geobacter species[J]. Energy & Environmental Science, 7(4): 1357-1361.
[18]	PAT-ESPADAS A M, RAZO-FLORES E, RANGEL-MENDEZ J R, et al., 2014. Direct and quinone-mediated palladium reduction by geobacter sulfurreducens: Mechanisms and modeling[J]. Environmental Science & Technology, 48(5): 2910-2919. DOI URL
[19]	SAINI G, CHAN C S, 2012. Near-neutral surface charge and hydrophilicity prevent mineral encrustation of Fe-oxidizing micro-organisms[J]. Geobiology, 11(2): 191-200. DOI URL
[20]	SCHEINOST A C, ROSSBERG A, VANTELON D, et al., 2006. Quantitative antimony speciation in shooting-range soils by EXAFS spectroscopy[J]. Geochimica et Cosmochimica Acta, 70(13): 3299-3312. DOI URL
[21]	SHI L D, WANG M, HAN Y L, et al., 2019. Multi-omics reveal various potential antimonate reductases from phylogenetically diverse microorganisms[J]. Applied Microbiology and Biotechnology, 103(21): 9119-9129. DOI
[22]	SUN W M, SUN X X, HAGGBLOM M M, et al., 2021. Identification of antimonate reducing bacteria and their potential metabolic traits by the combination of stable isotope probing and metagenomic-pangenomic analysis[J]. Environmental Science and Technology, 55(20): 13902-13912. DOI URL
[23]	THÖNY-MEYER L, 1997. Biogenesis of respiratory cytochromes in bacteria[J]. Microbiology and Molecular Biology Reviews, 61(3): 337-376.
[24]	TOLAR JOE G, LI S, AJO-FRANKLIN CAROLINE M, 2022. The differing roles of flavins and quinones in extracellular electron transfer in lactiplantibacillus plantarum[J]. Applied and Environmental Microbiology, 89(1): e01313-01322.
[25]	WANG L Y, YE L, JING C Y, 2020. Genetic identification of antimonate respiratory reductase in Shewanella sp. ANA-3[J]. Environmental Science & Technology, 54(21): 14107-14113. DOI URL
[26]	WANG L Y, YE L, YU Y Q, et al., 2018. Antimony redox biotransformation in the subsurface: Effect of indigenous Sb(V) respiring microbiota[J]. Environmental Science & Technology, 52(3): 1200-1207. DOI URL
[27]	WANG S S, XING Z H, CHEN G Y, et al., 2016. Cuboctahedral Sb₂O₃ mesocrystals organized from octahedral building blocks: More than self-similarity[J]. Crystal Growth & Design, 16(7): 3613-3617. DOI URL
[28]	WANG X M, WANG L, CHEN L, et al., 2022. AQDS activates extracellular synergistic biodetoxification of copper and selenite via altering the coordination environment of outer-membrane proteins[J]. Environmental Science & Technology, 56(19): 13786-13797. DOI URL
[29]	WANG X Q, LI F B, YUAN C L, et al., 2019. The translocation of antimony in soil-rice system with comparisons to arsenic: Alleviation of their accumulation in rice by simultaneous use of Fe(II) and NO₃^-[J]. Science of the Total Environment, 650(Part 1): 633-641. DOI URL
[30]	WU Y, LUO X, QIN B, et al., 2020. Enhanced current production by exogenous electron mediators via synergy of promoting biofilm formation and the electron shuttling process[J]. Environmental Science & Technology, 54(12): 7217-7225. DOI URL
[31]	YAMAMURA S, IIDA C, KOBAYASHI Y, et al., 2021. Production of two morphologically different antimony trioxides by a novel antimonate-reducing bacterium, Geobacter sp. SVR[J]. Journal of Hazardous Materials, 411: 125100. DOI URL
[32]	YANG Z R, HOSOKAWA H, SADAKANE T, et al., 2020. Isolation and characterization of facultative-anaerobic antimonate-reducing bacteria[J]. Microorganisms, 8(9): 1435. DOI URL
[33]	YING Z Y, CHEN H, HE Z, et al., 2022. Redox mediator-regulated microbial electrolysis cell to boost coulombic efficiency and degradation activity during gaseous chlorobenzene abatement[J]. Journal of Power Sources, 528: 231314.
[34]	YU H, YAN X Z, WENG W L, et al., 2022. Extracellular proteins of Desulfovibrio vulgaris as adsorbents and redox shuttles promote biomineralization of antimony[J]. J Hazard Mater, 426: 127795. DOI URL
[35]	YU Y S, CHEN J C, LI Y P, et al., 2021. Identification of a MarR subfamily that regulates arsenic resistance genes[J]. Applied and Environmental Microbiology, 87(24): e0158821. DOI URL
[36]	ZHANG Y D, BOYANOV M I, O’LOUGHLIN E J, et al., 2024. Reaction pathways and Sb(III) minerals formation during the reduction of Sb(V) by Rhodoferax ferrireducens strain YZ-1[J]. Journal of Hazardous Materials, 465: 133240. DOI URL
[37]	ZHOU J Z, WU C Y, PANG S, et al., 2022a. Dissimilatory and cytoplasmic antimonate reductions in a hydrogen-based membrane biofilm reactor[J]. Environmental Science & Technology, 56(20): 14808-14816. DOI URL
[38]	ZHOU J Z, WU C Y, PANG S, et al., 2022b. Dissimilatory and cytoplasmic antimonate reductions in a hydrogen-based membrane biofilm reactor[J]. Environmental Science & Technology, 56(20): 14808-14816. DOI URL
[39]	ZOTOV A V, SHIKINA N D, AKINFIEV N N, 2003. Thermodynamic properties of the Sb(III) hydroxide complex Sb(OH)_3(aq) at hydrothermal conditions[J]. Geochimica et Cosmochimica Acta, 67(10): 1821-1836. DOI URL
[40]	阳涅, 孙晓旭, 孔天乐, 等, 2023. 微生物群落对河流底泥中锑含量变化的响应[J]. 生态环境学报, 32(3): 609-618. DOI
	YANG N, SUN X X, KONG T L, et al.., 2023. Response of microbial communities to changes in antimony content in river sediments[J]. Journal of Ecology and Environment, 32(3): 609-618.

Microbial Mechanism of Electron Shuttle-mediated Antimony Reduction and Mineralization by Soil Microorganism

电子穿梭体介导土壤锑还原成矿的微生物机制

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