Mikrobiol. Z. 2020; 82(5):11-20.
Specificity of Lectins Labeled with Colloidal Gold to the Exopolymeric Matrix Carbohydrates
of the Sulfate-Reducing Bacteria Biofilm Formed on Steel
D.R. Abdulina, L.M. Purish, G.O. Iutynska
Zabolotny Institute of Microbiology and Virology, NAS of Ukraine
154 Akad. Zabolotny Str., Kyiv, 03143, Ukraine
The studies of the carbohydrate composition of the sulfate-reducing bacteria (SRB) biofilms formed on the steel surface, which are a factor of microbial corrosion, are significant. Since exopolymers synthesized by bacteria could activate corrosive processes. The aim of the study was to investigate the specificity of commercial lectins, labeled with colloidal gold to carbohydrates in the biofilm exopolymeric matrix produced by the corrosive-relevant SRB strains from man-caused ecotopes. Methods. Microbiological methods (obtaining of the SRB biofilms during cultivation in liquid Postgate B media under microaerophilic conditions), biochemical methods (lectin-binding analysis of 10 commercial lectins, labeled with colloidal gold), transmission electron microscopy using JEM-1400 JEOL. Results. It was shown using transmission electron microscopy that the binding of lectins with carbohydrates in the biofilm of the studied SRB strains occurred directly in the exopolymerіс matrix, as well as on the surfaces of bacterial cells, as seen by the presence of colloidal gold particles. For detection of the neutral carbohydrates (D-glucose and D-mannose) in the biofilm of almost all studied bacterial strains PSA lectin was the most specific. This lectin binding in biofilms of Desulfotomaculum sp. К1/3 and Desulfovibrio sp. 10 strains was higher in 90.8% and 94.4%, respectively, then for ConA lectin. The presence of fucose in the SRB biofilms was detected using LABA lectin, that showed specificity to the biofilm EPS of all the studied strains. LBA lectin was the most specific to N-аcetyl-D-galactosamine for determination of amino sugars in the biofilm. The amount of this lectin binding in D. vulgaris DSM644 biofilm was 30.3, 10.1 and 9.3 times higher than SBA, SNA and PNA lectins, respectively. STA, LVA and WGA lectins were used to detect the N-acetyl-Dglucosamine and sialic acid in the biofilm. WGA lectin showed specificity to N-acetyl-D-glucosamine in the biofilm of all the studied SRB; maximum number of bounded colloidal gold particles (175 particles/μm2) was found in the Desulfotomaculum sp. TC3 biofilm. STA lectin was interacted most actively with N-acetyl-D-glucosamine in Desulfotomaculum sp. TC3 and Desulfomicrobium sp. TC4 biofilms. The number of bounded colloidal gold particles was in 9.2 and 7.4 times higher, respectively, than using LVA lectin. The lowest binding of colloidal gold particles was observed for LVA lectin. Conclusions. It was identified the individual specificity of the 10 commercial lectins to the carbohydrates of biofilm matrix on the steel surface, produced by SRB. It was estimated that lectins with identical carbohydrates specificity had variation in binding to the biofilm carbohydrates of different SRB strains. Establishing of the lectin range selected for each culture lead to the reduction of the scope of studies and labor time in the researching of the peculiarities of exopolymeric matrix composition of biofilms formed by corrosiverelevant SRB.
Keywords: lectins, labeled with colloidal gold, exopolymeric matrix, sulfate-reducing bacteria, biofilm.
Full text (PDF, in English)
- Watnick P, Kolter R. Biofilm, city of microbes. J Bacteriol. 2000; 182(10):2675–79. https://doi.org/10.1128/JB.182.10.2675-2679.2000
- Davey ME, O’Toole GA. Microbial biofilms: from ecology to molecular genetics. Microbiol Mol Biol Rev. 2000; 64(4):847–67. https://doi.org/10.1128/MMBR.64.4.847-867.2000
- Vu B, Chen M, Crawford RJ, Ivanova EP. Bacterial extracellular polysaccharides involved in biofilm formation. Molecules. 2009; 14(7):2535−54. https://doi.org/10.3390/molecules14072535
- Flemming HC, Neu TR, Watnick D. The EPS matrix: The “House of biofilm cell”. J Bacteriol. 2007; 189(22):7945−47. https://doi.org/10.1128/JB.00858-07
- Lewandowski Z. Structure and function of bacterial biofilms. Biofilms: Recent Advances in Their Study and Control. L.V. Evans, ed. Harwood: Acad. Publishers; 2000. p. 21–79.
- Sutherland IW. Biofilm exopolysaccharides: a strong and sticky framework. Microbiology. 2001; 147(1):3–9. https://doi.org/10.1099/00221287-147-1-3
- Hamilton WA. Microbially influenced corrosion as a model system for the study of metal microbe interactions: a unifying electron transfer hypothesis. Biofouling. 2003; 19(1):65−76. https://doi.org/10.1080/0892701021000041078
- Beech IW, Summer J. Biocorrosion: towards understanding interactions between biofilms and metals. Current Opinion Biotechnol. 2004; 15(3):181−6. https://doi.org/10.1016/j.copbio.2004.05.001
- Lahtin V.M. [Lektiny v issledovanii uglevodnoi chasti glycoproteinov I drugih prirodnyh glicoconjugantov]. Biochemistry (Biokhimiya). 1995; 60(2):187–217. Russian.
- Neu TR, Swerhone DW, Lawrence JR. Assessment of lectin-binding analysis for in situ detection of glycoconjugates in biofilm systems. Microbiology. 2001; 147:299-313. https://doi.org/10.1099/00221287-147-2-299
- Neu TR, Manz B, Volke F, Dynes JJ, et al. Advanced imaging techniques for assessment of structure, composition and function in biofilm systems. FEMS Microbiol Ecol. 2010; 72(1):1–21. https://doi.org/10.1111/j.1574-6941.2010.00837.x
- Lawrence JR, Swerhone GDW, Leppard GG, Araki T, Zhang X, West MM, Hitchcock AP. Scanning transmission X-ray, laser scanning, and transmission electron microscopy mapping of exopolymers matrix of microbial biofilms. Appl Environ Microbiol. 2003; 69(9):5543−54. https://doi.org/10.1128/AEM.69.9.5543-5554.2003
- Zippel B, Neu TRB. Characterization of glycoconjugates of extracellular polymeric substances in tufa-associated biofilms by using fluorescence lectin-binding analysis. Appl Environ Microbiol. 2011; 77(2):505–16. https://doi.org/10.1128/AEM.01660-10
- Lawrence JR, Swerhone GDW, Neu TR. In situ evidence for microdomains in the polymer matrix of bacterial microcolonies. Can J Microbiol. 2007; 53(3):450–458. https://doi.org/10.1139/W06-146
- Purish LM, Asaulenko LG, Abdulina DR, Voychuk SI, Iutynskaya GA. Lectin-binding analysis of the biofilm exopolymeric matrix carbohydrate composition of corrosion-aggressive bacteria. Appl Biochem Microbiol. 2013; 49(5):458–63. https://doi.org/10.1134/S0003683813050104
- Purish LM, Asaulenko LG, Abdulina DR, Vasil’ev VN, Iutinskaya GA. Role of polymer complexes in the formation of biofilms by corrosive bacteria on steel surfaces. Appl Biochem Microbiol. 2012; 48(3):262–9. https://doi.org/10.1134/S0003683812030118
- Asaulenko LG, Abdulina DR, Purish LM. [Taxonomic position of certain representatives of sulphate-reducing corrosive microbial community]. Mikrobiol Z. 2010; 72(4):3–10. Ukrainian.
- Purish LM, Asaulenko LG, Abdulina DR, Iutinskaia GA. [Biodiversity of sulfate-reducing bacteria growing on objects of heating systems]. Mikrobiol Z. 2014; 76(3):11–17. Russian.
- Abdulina DR, Purish LM, Iutynska GO. Microbial communities and sulfate-reducing bacteria in soils near main-gas pipeline. Mikrobiol Z. 2018; 80(5):3–14. https://doi.org/10.15407/microbiolj80.05.003
- Elgavish S, Shaanan B. Lectin-carbohydrate interactions: different folds, common recognition principles. Trends Biochem Sci. 1997; 22(12):462–67. https://doi.org/10.1016/S0968-0004(97)01146-8
- Beech I, Zinkevich V, Tapper R, Gubner R, Avci R. Study of interaction of sulphate-reducing bacteria exopolymers with iron using X-ray photoelectron spectroscopy and time-of-flight secondary ionisation mass spectrometry. J Microbiol Methods. 1999; 36(1–2):3–10. https://doi.org/10.1016/S0167-7012(99)00005-6
- Liener IE, Sharon N, Goldstein IJ. The Lectins: properties, functions and applications in biology and medicine. Orlando: Acad Press; 1986.
- Weiss WI, Drickamer K. Structural basis of lectin-carbohydrate recognition. Ann Rev Biochem. 1996; 65:441–473. https://doi.org/10.1146/annurev.bi.65.070196.002301
- Castro L, Zhang R, Munoz JA, Gonzalez F, Blazquez ML, Sand W, Ballester A. Characterization of exopolymeric substances (EPS) produced by Aeromonas hydrophila under reducing conditions. Biofouling. 2014; 30(4):501–11. https://doi.org/10.1080/08927014.2014.892586
- Neu TR, Lawrence JR. Advanced techniques for in situ analysis of the biofilm matrix (structure, composition, dynamics) by means of laser scanning microscopy. In Microbial Biofilms: Methods and Protocols, Methods. In Molecular Biology. G. Donelli, ed. New York, USA: Springer; 2014. p. 43–64. https://doi.org/10.1007/978-1-4939-0467-9_4
- Neu TR, Kuhlicke U. Fluorescence lectin bar-coding of glycoconjugates in the extracellular matrix of biofilm and bio aggregate forming microorganisms. Microorganisms. 2017; 5(1):1–13. https://doi.org/10.3390/microorganisms5010005
- Bennke CM, Neu TR, Fuchs BM, Amann R. Mapping glycoconjugate- mediated interactions of marine Bacteroidetes with diatoms. Syst Appl Microbiol. 2013; 36:417–25. https://doi.org/10.1016/j.syapm.2013.05.002
- Zhang RY, Neu TR, Bellenberg S, et al. Use of lectins to in situ visualize glycoconjugates of extracellular polymeric substances in acidophilic archaeal biofilm. Microb Biotechnol. 2015; 8(3):448–61. https://doi.org/10.1111/1751-7915.12188