Distribution and content changes of extracellular polymeric substance and iron ions on the pyrite surface during bioleaching

Abstract

Microbial-mineral interface is the main biochemical reaction place of bioleaching, and the mechanism of interface interaction is the key to elucidating the behavior of microbial leaching. In this paper, bioleaching experiments and biofilm colonization experiments of pyrite grains and pyrite slices were conducted by three moderately thermophilic mixed bacteria, Acidithiobacillus caldus, Leptospirillum ferriphilum and Sulfobacillus thermosulfidooxidans, respectively. The distribution and content changes of interfacial extracellular polymeric substance (EPS) and iron ions during bioleaching were evaluated based on a highly selective and sensitive metal fluorescence probe combined with confocal laser scanning microscopy (CLSM), scanning electron microscopy (SEM) and physical extraction methods. The results show that EPS was laid flat on the surface of pyrite slices to form sheet biofilms, mainly in the early and middle stages of bioleaching, reaching 17.21 mg/g on the 15th day, and the main component was extracellular protein. At the early stage of bioleaching, the surface of pyrite was mainly dominated by Fe²⁺ of 0.75 mg/g, while Fe³⁺ was only 0.08 mg/g. With the increase of leaching time, a large amount of Fe³⁺ (11.97 mg/g) was enriched at the microbe-mineral interface.

Keywords

bioleaching / pyrite / mixed culture / interface / biofilm / metal ion / extracellular polymeric substance / confocal laser scanning microscopy

References

[1] RohwerderT, GehrkeT, KinzlerK, et al. . Bioleaching review, part A [J]. Applied Microbiology and Biotechnology, 2003, 63(3): 239-248

[2] RobertoF F, SchippersA. Progress in bioleaching: Part B. Applications of microbial processes by the minerals industries [J]. Applied Microbiology and Biotechnology, 2022, 106(18): 5913-5928

[3] WuX, WuX, DengF, et al. . Comparison of bioleaching of chalcopyrite concentrates with mixed culture after cryopreservation with PEG-2000 in liquid nitrogen [J]. Journal of Central South University, 2020, 27(5): 1386-1394

[4] YinL, YangH, LuL, et al. . Interfacial alteration of pyrite caused by bioleaching [J]. Hydrometallurgy, 2020, 195: 105356

[5] MehrabaniJ V, ShafaeiS Z, NoaparastM, et al. . Bioleaching of different pyrites and sphalerite in the presence of graphite [J]. Geomicrobiology Journal, 2017, 34(2): 97-108

[6] AiC, LiangY, QiuG, et al. . Bioleaching of low-grade copper sulfide ore by extremely thermoacidophilic consortia at 70 °C in column reactors [J]. Journal of Central South University, 2020, 27(5): 1404-1415

[7] ZhuW, XiaJ, YangY, et al. . Sulfur oxidation activities of pure and mixed thermophiles and sulfur speciation in bioleaching of chalcopyrite [J]. Bioresource Technology, 2011, 102(4): 3877-3882

[8] LeahyM J, DavidsonM R, SchwarzM P. A model for heap bioleaching of chalcocite with heat balance: Mesophiles and moderate thermophiles [J]. Hydrometallurgy, 2007, 85(1): 24-41

[9] D’HuguesP, FoucherS, Gallé-CavalloniP, et al. . Continuous bioleaching of chalcopyrite using a novel extremely thermophilic mixed culture [J]. International Journal of Mineral Processing, 2002, 66(1–4): 107-119

[10] BehradV A. A comparison of bioleaching ability of mesophilic and moderately thermophilic culture on copper bioleaching from flotation concentrate and smelter dust [J]. International Journal of Mineral Processing, 2011, 101(1–4): 94-99

[11] ShenL, ChengJ, WangJ, et al. . Comparison of extraction methods for extracellular polymeric substances (EPS) and dynamic characterization of EPS from sessile microorganisms during pyrite bioleaching [J]. Journal of Environmental Chemical Engineering, 2022, 10(3): 107922

[12] XiaL, DaiS, YinC, et al. . Comparison of bioleaching behaviors of different compositional sphalerite using Leptospirillum ferriphilum, Acidithiobacillus ferrooxidans and Acidithiobacillus caldus [J]. Journal of Industrial Microbiology & Biotechnology, 2009, 366845-851

[13] LiQ, ZhuJ, LiS, et al. . Interactions between cells of Sulfobacillus thermosulfidooxidans and Leptospirillum ferriphilum during pyrite bioleaching [J]. Frontiers in Microbiology, 2020, 1144

[14] VardanyanN, BadalyanH, MarkosyanL, et al. . Newly isolated Acidithiobacillus sp. ksh from kashen copper ore: Peculiarities of EPS and colloidal exopolysaccharide [J]. Frontiers in Microbiology, 2020, 111802

[15] VeraM, SchippersA, SandW. Progress in bioleaching: Fundamentals and mechanisms of bacterial metal sulfide oxidation—Part A [J]. Applied Microbiology and Biotechnology, 2013, 97(17): 7529-7541

[16] ZhangR, NeuT R, BlanchardV, et al. . Biofilm dynamics and EPS production of a thermoacidophilic bioleaching archaeon [J]. New Biotechnology, 2019, 5121-30

[17] FlemmingH C, WingenderJ, SzewzykU, et al. . Biofilms: An emergent form of bacterial life [J]. Nature Reviews Microbiology, 2016, 14(9): 563-575

[18] VuB, ChenM, CrawfordR J, et al. . Bacterial extracellular polysaccharides involved in biofilm formation [J]. Molecules (Basel, Switzerland), 2009, 14(7): 2535-2554

[19] FlemmingH C, WingenderJ. The biofilm matrix [J]. Nature Reviews Microbiology, 2010, 8(9): 623-633

[20] LiQ, SandW. Mechanical and chemical studies on EPS from Sulfobacillus thermosulfidooxidans: From planktonic to biofilm cells [J]. Colloids and Surfaces B: Biointerfaces, 2017, 153: 34-40

[21] HeZ, YangY, ZhouS, et al. . Effect of pyrite, elemental sulfur and ferrous ions on EPS production by metal sulfide bioleaching microbes [J]. Transactions of Nonferrous Metals Society of China, 2014, 24(4): 1171-1178

[22] LiQ, SandW, ZhangR. Enhancement of biofilm formation on pyrite by Sulfobacillus thermosulfidooxidans [J]. Minerals, 2016, 6(3): 71

[23] ZengW, CaiY, HouC, et al. . Influence diversity of extracellular DNA on bioleaching chalcopyrite and pyrite by Sulfobacillus thermosulfidooxidans ST [J]. Journal of Central South University, 2020, 2751466-1476

[24] KinzlerK, GehrkeT, TelegdiJ, et al. . Bioleaching-A result of interfacial processes caused by extracellular polymeric substances (EPS) [J]. Hydrometallurgy, 2003, 71(1–2): 83-88

[25] ZhangR, DuanJ, XuD, et al. . Bioleaching and biocorrosion: Advances in interfacial processes [J]. Frontiers in Microbiology, 2021, 12: 653029

[26] VilinskaA, RaoK H. Surface thermodynamics and extended DLVO theory of Acidithiobacillus ferrooxidans cells adhesion on pyrite and chalcopyrite [J]. The Open Colloid Science Journal, 2009, 2(1): 1-14

[27] VardanyanA, VardanyanN, KhachatryanA, et al. . Adhesion to mineral surfaces by cells of leptospirillum, acidithiobacillus and sulfobacillus from Armenian sulfide ores [J]. Minerals, 2019, 9(2): 69

[28] LiQ, WangQ, ZhuJ, et al. . Effect of extracellular polymeric substances on surface properties and attachment behavior of Acidithiobacillus ferrooxidans [J]. Minerals, 2016, 64100

[29] YuR, TanJ, GuG, et al. . Mechanism of bioleaching chalcopyrite by Acidithiobacillus ferrooxidans in agar-simulated extracellular polymeric substances media [J]. Journal of Central South University of Technology, 2010, 17156-61

[30] SandW, GehrkeT. Extracellular polymeric substances mediate bioleaching/biocorrosion via interfacial processes involving iron(III) ions and acidophilic bacteria [J]. Research in Microbiology, 2006, 157(1): 49-56

[31] GieseE C. Evidences of EPS-iron (III) ions interactions on bioleaching process mini-review: The key to improve performance [J]. Orbital: the Electronic Journal of Chemistry, 2019, 11(3): 200-204

[32] FowlerT A, CrundwellF K. Leaching of zinc sulfide by thiobacillus ferrooxidans: Experiments with a controlled redox potential indicate no direct bacterial mechanism [J]. Applied and Environmental Microbiology, 1998, 64(10): 3570-3575

[33] ZhangR, BellenbergS, CastroL, et al. . Colonization and biofilm formation of the extremely acidophilic archaeon Ferroplasma acidiphilum [J]. Hydrometallurgy, 2014, 150245-252

[34] ZengW, QiuG, ZhouH, et al. . Characterization of extracellular polymeric substances extracted during the bioleaching of chalcopyrite concentrate [J]. Hydrometallurgy, 2010, 100(3–4): 177-180

[35] ZengW, TanS, ChenM, et al. . Detection and analysis of attached microorganisms on the mineral surface during bioleaching of pure chalcopyrite with moderate thermophiles [J]. Hydrometallurgy, 2011, 106(1–2): 46-50

[36] SheM, YangZ, YinB, et al. . A novel rhodamine-based fluorescent and colorimetric “off-on” chemosensor and investigation of the recognizing behavior towards Fe³⁺ [J]. Dyes and Pigments, 2012, 92(3): 1337-1343

[37] ZhangX, BishopP L, KinkleB K. Comparison of extraction methods for quantifying extracellular polymers in biofilms [J]. Water Science and Technology, 1999, 39(7): 211-218

[38] PengT, ZhouD, LiuX, et al. . Enrichment of ferric iron on mineral surface during bioleaching of chalcopyrite [J]. Transactions of Nonferrous Metals Society of China, 2016, 26(2): 544-550

[39] SaterlayA J, HongQ, ComptonR G, et al. . Ultrasonically enhanced leaching: Removal and destruction of cyanide and other ions from used carbon cathodes [J]. Ultrasonics Sonochemistry, 2000, 7(1): 1-6

[40] AfzalG M, OkibeN, BarrieJ D. Attachment of acidophilic bacteria to solid surfaces: The significance of species and strain variations [J]. Hydrometallurgy, 2007, 85(2–4): 72-80

[41] CowanD H, JahromiF G, GhahremanA. Atmospheric oxidation of pyrite with a novel catalyst and ultra-high elemental sulphur yield [J]. Hydrometallurgy, 2017, 173: 156-169

[42] DescostesM, VitorgeP, BeaucaireC. Pyrite dissolution in acidic media [J]. Geochimica et Cosmochimica Acta, 2004, 68(22): 4559-4569

[43] RodríguezY, BallesterA, BlázquezM L, et al. . New information on the pyrite bioleaching mechanism at low and high temperature [J]. Hydrometallurgy, 2003, 71(1–2): 37-46

[44] KocamanA T, CemekM, EdwardsK J. Kinetics of pyrite, pyrrhotite, and chalcopyrite dissolution by Acidithiobacillus ferrooxidans [J]. Canadian Journal of Microbiology, 2016, 62(8): 629-642

[45] GehrkeT, TelegdiJ, ThierryD, et al. . Importance of extracellular polymeric substances from thiobacillus ferrooxidans for bioleaching [J]. Applied and Environmental Microbiology, 1998, 64(7): 2743-2747

[46] CaoL, HuangZ, SunX, et al. . Comparison of leaching of bornite from different regions mediated by mixed moderately thermophilic bacteria [J]. Journal of Central South University, 2020, 27(5): 1373-1385

[47] BramhachariP V, NagarajuG P. Extracellular polysaccharide production by bacteria as a mechanism of toxic heavy metal biosorption and biosequestration in the marine environment [M]. Marine Pollution and Microbial Remediation, 2016, Singapore, Springer Singapore: 6785

[48] TianX, ShenZ, HanZ, et al. . The effect of extracellular polymeric substances on exogenous highly toxic compounds in biological wastewater treatment: An overview [J]. Bioresource Technology Reports, 2019, 5: 28-42

[49] HaoL, GuoY, ByrneJ M, et al. . Binding of heavy metal ions in aggregates of microbial cells, EPS and biogenic iron minerals measured in-situ using metal- and glycoconjugates-specific fluorophores [J]. Geochimica et Cosmochimica Acta, 2016, 180: 66-96

[50] HaoL, LiJ, KapplerA, et al. . Mapping of heavy metal ion sorption to cell-extracellular polymeric substance-mineral aggregates by using metal-selective fluorescent probes and confocal laser scanning microscopy [J]. Applied and Environmental Microbiology, 2013, 79(21): 6524-6534

[51] MitsunobuS, ZhuM, TakeichiY, et al. . Direct detection of Fe(II) in extracellular polymeric substances (EPS) at the mineral-microbe interface in bacterial pyrite leaching [J]. Microbes and Environments, 2016, 31(1): 63-69

[52] CastroL, ZhangR, MuñozJ A, et al. . Characterization of exopolymeric substances (EPS) produced by aeromonas hydrophila under reducing conditions [J]. Biofouling, 2014, 30(4): 501-511

[53] SandW, GehrkeT, JozsaP G, et al. . (Bio)chemistry of bacterial leaching—Direct vs. indirect bioleaching [J]. Hydrometallurgy, 2001, 59(2–3): 159-175

[54] YuR, ZhongD, MiaoL, et al. . Relationship and effect of redox potential, jarosites and extracellular polymeric substances in bioleaching chalcopyrite by acidithiobacillus ferrooxidans [J]. Transactions of Nonferrous Metals Society of China, 2011, 21(7): 1634-1640

[55] YuR, LiuJ, ChenA, et al. . Interaction mechanism of Cu²⁺, Fe³⁺ ions and extracellular polymeric substances during bioleaching chalcopyrite by Acidithiobacillus ferrooxidans ATCC2370 [J]. Transactions of Nonferrous Metals Society of China, 2013, 23(1): 231-236