Characterization of Biofilm Extracts from Two Marine Bacteria
Source abstract
Abstract
In the marine environment, biofilm formation is an important lifestyle for microorganisms. A biofilm is comprised of cells embedded in an extracellular matrix that holds them close together and keeps the biofilm attached to the colonized surface. This predominant lifestyle and its main regulation pathway, namely quorum-sensing (QS), have been shown to induce specific bioactive metabolites. In this study, we investigated the biofilm formation by two marine bacteria belonging to the Vibrio species to discover potentially innovative bioactive compounds. We proposed a protocol to isolate biofilm extracts, to analyze their biochemical composition, and to compare them to planktonic cell extracts. Cells were grown attached to a plastic surface; extracts were prepared in water, NaOH, or in ethyl acetate and analyzed. Extracellular matrix components featured carbohydrates, proteins, lipids, and low amount of DNA. Carbohydrates appeared to be the main constituent of biofilm but also of the planktonic cell supernatant. Moreover, antimicrobial and QS-signaling activities were evidenced in extracts.
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Research context
OpenAlex topics: Bacterial biofilms and quorum sensing · Oral microbiology and periodontitis research · Vibrio bacteria research studies
SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.
Affiliations
Ifremer, Atlantique Center, Microbial Ecosystems and Marine Molecules for the Biotechnologies, 44311 Rue de l’Ile d’Yeu, Nantes CEDEX 3, France
Ifremer, Bretagne Center, Health, Environment and Microbiology, 1625 Route de Sainte-Anne, 29280 Plouzané, France
Funding
Institut Français de Recherche pour l'Exploitation de la Mer
References (83)
Show source-provided references
1. Bacteria and archaea on Earth and their abundance in biofilms10.1038/s41579-019-0158-9 ↗
2. Neu, T.R., and Lawrence, J.R. (2010). Chapter 37—Extracellular polymeric substances in microbial biofilms. Microbial Glycobiology, Academic Press.10.1016/B978-0-12-374546-0.00037-7 ↗
3. The biofilm matrix10.1038/nrmicro2415 ↗
4. Marine Biofilms: A successful microbial strategy with economic implications10.3389/fmars.2018.00126 ↗
5. Cyclic-di-GMP Regulates Extracellular Polysaccharide Production, Biofilm Formation, and Rugose Colony Development by Vibrio vulnificus10.1128/AEM.00176-08 ↗
6. Quorum sensing and bacterial biofilms10.1039/b804469b ↗
7. Biofilm Formation and Heat Stress Induce Pyomelanin Production in Deep-Sea Pseudoalteromonas sp. SM991310.3389/fmicb.2017.01822 ↗
8. Induced Biofilm Cultivation Enhances Riboflavin Production by an Intertidally Derived Candida famata10.1007/s12010-012-9626-7 ↗
9. Glycolipid biosurfactants: Potential related biomedical and biotechnological applications10.1016/j.carres.2015.07.016 ↗
10. Quorum sensing as a potential target for increased production of rhamnolipid biosurfactant in Burkholderia thailandensis E26410.1007/s00253-019-09942-5 ↗
11. Production of the antifungal compound pyrrolnitrin is quorum sensing-regulated in members of theBurkholderia cepaciacomplex10.1111/j.1462-2920.2009.01870.x ↗
12. Relevance of microbial extracellular polymeric substances (EPSs)—Part I: Structural and ecological aspects10.2166/wst.2001.0326 ↗
13. Characterization of extracellular polymers synthesized by tropical intertidal biofilm bacteria10.1111/j.1365-2672.2006.03085.x ↗
14. Bacterial Extracellular Polysaccharides in Biofilm Formation and Function10.1128/microbiolspec.MB-0011-2014 ↗
15. Biofilm exopolysaccharides: A strong and sticky framework10.1099/00221287-147-1-3 ↗
16. Ostapska, H., Howell, P.L., and Sheppard, D.C. (2018). Deacetylated microbial biofilm exopolysaccharides: It pays to be positive. PLoS Pathog., 14.10.1371/journal.ppat.1007411 ↗
17. Kjelleberg, S., and Givskov, M. (2007). The biofilm matrix—A sticky framework. The Biofilm Mode of Life, Horizon Press.
18. Marine Polysaccharides: A Source of Bioactive Molecules for Cell Therapy and Tissue Engineering10.3390/md9091664 ↗
19. Xanthobacter-dominated biofilm as a novel source for high-value rhamnose10.1007/s00253-019-09765-4 ↗
20. Production and biotechnological potential of extracellular polymeric substances from sponge-associated antarctic bacteria10.1128/AEM.01624-17 ↗
21. Establishing a Role for Bacterial Cellulose in Environmental Interactions: Lessons Learned from Diverse Biofilm-Producing Proteobacteria10.3389/fmicb.2015.01282 ↗
22. Pseudomonas aeruginosa lectin LecB is located in the outer membrane and is involved in biofilm formation10.1099/mic.0.27701-0 ↗
23. Bap: A family of surface proteins involved in biofilm formation10.1016/j.resmic.2005.11.003 ↗
24. Exoenzyme accumulation in epilithic biofilms10.1007/BF00017497 ↗
25. Extracellular nucleases and extracellular DNA play important roles in Vibrio cholerae biofilm formation10.1111/j.1365-2958.2011.07867.x ↗
26. Extracellular DNA and lipoteichoic acids interact with exopolysaccharides in the extracellular matrix of Streptococcus mutansbiofilms10.1080/08927014.2017.1361412 ↗
27. Identification of Extracellular DNA-Binding Proteins in the Biofilm Matrix10.1128/mBio.01137-19 ↗
28. Gene transfer occurs with enhanced efficiency in biofilms and induces enhanced stabilisation of the biofilm structure10.1016/S0958-1669(03)00036-3 ↗
29. Sensor kinase RscS induces the production of antigenically distinct outer membrane vesicles that depend on the symbiosis polysaccharide locus in Vibrio fischeri10.1128/JB.05926-11 ↗
30. Microbial Extracellular Polymeric Substances (EPSs) in Ocean Systems10.3389/fmicb.2017.00922 ↗
31. Importance of Extracellular Polymeric Substances from Thiobacillus ferrooxidans for Bioleaching10.1128/AEM.64.7.2743-2747.1998 ↗
32. Growth of Mycobacterium tuberculosis biofilms containing free mycolic acids and harbouring drug-tolerant bacteria10.1111/j.1365-2958.2008.06274.x ↗
33. Rhamnolipid Surfactant Production Affects Biofilm Architecture in Pseudomonas aeruginosa PAO110.1128/JB.185.3.1027-1036.2003 ↗
34. Extracellular polymeric substances of biofilms: Suffering from an identity crisis10.1016/j.watres.2018.11.020 ↗
35. Characterization of extracellular polymeric matrix, and treatment of Fusobacterium nucleatum and Porphyromonas gingivalis biofilms with DNase I and proteinase K
36. Vibrio cholerae O1 Strain TSI-4 Produces the Exopolysaccharide Materials That Determine Colony Morphology, Stress Resistance, and Biofilm Formation10.1128/AEM.64.10.3648-3655.1998 ↗
37. Ramirez-Mora, T., Retana-Lobo, C., and Valle-Bourrouet, G. (2018). Biochemical characterization of extracellular polymeric substances from endodontic biofilms. PLoS ONE, 13.10.1371/journal.pone.0204081 ↗
38. Comparison of two chemical extraction methods for proteins and polysaccharides of Spirogyra fluviatilis in extracellular polymeric substances
39. Characterization and anti-biofilm activity of extracellular polymeric substances produced by the marine biofilm-forming bacteriumPseudoalteromonas ulvaestrain TC1410.1080/08927014.2016.1164845 ↗
40. Vibrio diabolicus sp. nov., a New Polysaccharide-Secreting Organism Isolated from a Deep-Sea Hydrothermal Vent Polychaete Annelid, Alvinella pompejana10.1099/00207713-47-4-989 ↗
41. Deep-sea hydrothermal vents: A new source of innovative bacterial exopolysaccharides of biotechnological interest?10.1038/sj.jim.7000298 ↗
42. Delbarre-Ladrat, C., Salas, M.L., Sinquin, C., Zykwinska, A., and Colliec-Jouault, S. (2017). Bioprospecting for Exopolysaccharides from Deep-Sea Hydrothermal Vent Bacteria: Relationship between Bacterial Diversity and Chemical Diversity. Microorganisms, 5.10.3390/microorganisms5030063 ↗
43. BioEdit: A user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT
44. MEGA7: Molecular Evolutionary Genetics Analysis version 7.0 for bigger datasets10.1093/molbev/msw054 ↗
45. The neighbor-joining method: A new method for reconstructing phylogenetic trees
46. Confidence Limits on Phylogenies: An Approach Using the Bootstrap10.2307/2408678 ↗
47. Jukes, T.H., and Cantor, C.R. (1969). Evolution of Protein Molecules. Mammalian Protein Metabolism, Academic Press.10.1016/B978-1-4832-3211-9.50009-7 ↗
48. Measurement of protein using bicinchoninic acid10.1016/0003-2697(85)90442-7 ↗
49. Über den Gehalt der wichtigsten Proteine der Nahrungsmittel an Kohlenhydrat und über ein kolorimetrisches Verfahren zur quantitativen Bestimmung von stickstoff-freiem Zucker im Eiweiß
50. The carbohydrate complex of the serum proteins improved method for isolation and re-determination of structure. Isolation of glucosaminodimannose from proteins of ox blood10.1042/bj0251062 ↗
51. A rapid method of total lipid extraction and purification10.1139/y59-099 ↗
52. Axelsson, M., and Gentili, F. (2014). A Single-Step Method for Rapid Extraction of Total Lipids from Green Microalgae. PLoS ONE, 9.10.1371/journal.pone.0089643 ↗
53. Improved determination of total serum lipids by the sulfo-phospho-vanillin reaction10.1093/clinchem/18.7.673 ↗
54. Zykwinska, A., Marchand, L., Bonnetot, S., Sinquin, C., Colliec-Jouault, S., and Delbarre-Ladrat, C. (2019). Deep-sea Hydrothermal Vent Bacteria as a Source of Glycosaminoglycan-Mimetic Exopolysaccharides. Molecules, 24.10.3390/molecules24091703 ↗
55. Assessment of biochemical methods to detect enzymatic depolymerization of polysaccharides10.1016/j.carbpol.2008.10.022 ↗
56. Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T410.1038/227680a0 ↗
57. Characterization by gas-liquid chromatography mass spectrometry of permethylsilylglycosides obtained in the methanolysis of glycoproteins and glycolipids10.1042/bj1510491 ↗
58. Chaplin, M.F., and Kennedy, J.K. (1986). Glycoproteins. Carbohydrate Analysis: A Practical Approach, IRL Press. [2nd ed.].
59. Techniques sélectives pour la recherche systématique des germes antibiotiques
60. Updating the Vibrio clades defined by multilocus sequence phylogeny: Proposal of eight new clades, and the description of Vibrio tritonius sp. nov10.3389/fmicb.2013.00414 ↗
61. New Vibrio species associated to molluscan microbiota: A review
62. Unusual Glycosaminoglycans from a Deep Sea Hydrothermal Bacterium Improve Fibrillar Collagen Structuring and Fibroblast Activities in Engineered Connective Tissues10.3390/md11041351 ↗
63. Genome sequence of Vibrio diabolicus and identification of the exopolysaccharide HE800 biosynthesis locus10.1007/s00253-014-6086-8 ↗
64. Bacterial exopolysaccharide mediated heavy metal removal: A review on biosynthesis, mechanism and remediation strategies10.1016/j.btre.2016.12.006 ↗
65. VpsR and cyclic di-GMP together drive transcription initiation to activate biofilm formation in Vibrio cholerae10.1093/nar/gky606 ↗
66. Yildiz, F., Fong, J., Sadovskaya, I., Grard, T., and Vinogradov, E. (2014). Structural Characterization of the Extracellular Polysaccharide from Vibrio cholerae O1 El-Tor. PLoS ONE, 9.10.1371/journal.pone.0086751 ↗
67. Structural studies of the main exopolysaccharide produced by the deep-sea bacterium Alteromonas infernus10.1016/j.carres.2004.07.021 ↗
68. Structure of the exopolysaccharide of Vibriodiabolicus isolated from a deep-sea hydrothermal vent10.1016/S0008-6215(99)00214-1 ↗
69. A color-code for glycosaminoglycans identification by means of polyacrylamide gel electrophoresis stained with the cationic carbocyanine dye Stains-all10.1002/elps.201700391 ↗
70. Roles of the Structural Symbiosis Polysaccharide (syp) Genes in Host Colonization, Biofilm Formation, and Polysaccharide Biosynthesis in Vibrio fischeri10.1128/JB.00707-12 ↗
71. Lebellenger, L., Verrez-Bagnis, V., Passerini, D., and Delbarre-Ladrat, C. (2018). Comparative genomics reveals a widespread distribution of an exopolysaccharide biosynthesis gene cluster among Vibrionaceae. BMC Res. Notes, 11.10.1186/s13104-018-3214-z ↗
72. Isolation and physico-chemical characterisation of extracellular polymeric substances produced by the marine bacterium Vibrio parahaemolyticus10.1080/08927014.2011.562605 ↗
73. Identification of a c-di-GMP-Regulated Polysaccharide Locus Governing Stress Resistance and Biofilm and Rugose Colony Formation in Vibrio vulnificus10.1128/IAI.01188-09 ↗
74. The surprisingly diverse ways that prokaryotes move10.1038/nrmicro1900 ↗
75. Identification of genes induced in Vibrio cholerae in a dynamic biofilm system10.1016/j.ijmm.2014.05.011 ↗
76. Identification of capsule, biofilm, lateral flagellum, and type IV pili in Vibrio mimicus strains10.1016/j.micpath.2014.09.012 ↗
77. Production of Bioactive Secondary Metabolites by Marine Vibrionaceae10.3390/md9091440 ↗
78. Antibacterial Compounds from Marine Vibrionaceae Isolated on a Global Expedition10.3390/md8122946 ↗
79. Quorum Sensing in the Squid-Vibrio Symbiosis10.3390/ijms140816386 ↗
80. Activity, Abundance, and Localization of Quorum Sensing Receptors in Vibrio harveyi10.3389/fmicb.2017.00634 ↗
81. Three Parallel Quorum-Sensing Systems Regulate Gene Expression in Vibrio harveyi10.1128/JB.186.20.6902-6914.2004 ↗
82. Casillo, A., Lanzetta, R., Parrilli, M., and Corsaro, M.M. (2018). Exopolysaccharides from Marine and Marine Extremophilic Bacteria: Structures, Properties, Ecological Roles and Applications. Mar. Drugs, 16.10.3390/md16020069 ↗
83. Prospecting for new bacterial metabolites: A glossary of approaches for inducing, activating and upregulating the biosynthesis of bacterial cryptic or silent natural products10.1039/C5NP00111K ↗