Phylogenetics and antibacterial properties of exopolysaccharides from marine bacteria isolated from Mauritius seawater
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The marine environment harbours diverse bacterial species which can be exploited for the production of valuable compounds such as exopolysaccharides (EPS) which hold promises for biotechnological applications. The coastal waters of Mauritius is a relatively underexplored marine environment and in this study, isolated bacterial species were tested for the production of EPS exhibiting antibacterial properties against human bacterial pathogens from the genera Acinetobacter, Bacillus, Campylobacter, Enterobacter, Enterococcus, Escherichia, Proteus, Pseudomonas, Salmonella, Streptococcus and Staphylococcus.
Bacteria were first isolated from seawater samples. Using the disc diffusion method, their EPS were tested for antibacterial effects through two screenings, with each involving a different set of arbitrarily chosen group of pathogens. The microorganisms producing antibacterial EPS were subsequently identified by morphological, biochemical and 16S rRNA-based phylogenetic analyses. Those EPS exhibiting broadest antibacterial activities were eventually characterised by Fourier-transform infrared spectroscopy (FTIR) and thin-layer chromatography (TLC).
Eight EPS were found to display antibacterial effects against more than half of the pathogens and the microorganisms producing them were identified as Bacillus, Halomonas, Psychrobacter and Alcaligenes species. However, only two of these EPS were found to be the most active, with their MIC values ranging between 62.5 and 500 μg/ml. FTIR and TLC analyses revealed the presence of carboxyl, hydroxyl and amide as well as sulphate for the EPS, with glucose or fructose being the main sugar.
The results suggest that Mauritius seawater can be a source of biotechnologically useful microorganisms, producing EPS having potential as antimicrobial agents. DNA sequence data also suggest possible novel bacterial species.
KeywordsAntimicrobial Bacteria Exopolysaccharides Mauritius Phylogeny Seawater
The authors wish to thank the Faculty of Agriculture, Faculty of Science, CBBR and University of Mauritius for supporting this study. The support of the technical staff of the Department of Agriculture & Food Science as well as the Department of Chemistry is gratefully acknowledged.
This work was supported by a grant from the University of Mauritius (grant number Q0117).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
Research involving human participants and/or animals
This article does not contain any studies with human participants or animals performed by any of the authors.
- Balzaretti S, Taverniti V, Guglielmetti S, Fiore W, Minuzzo M, Ngo HN, Ngere JB, Sadiq S, Humphreys PN, Laws AP (2017) A novel rhamnose-rich heteroexopolysaccharide isolated from Lactobacillus paracasei DG activates THP-1 human monocytic cells. Appl Environ Microbiol 83(3):e02702–e02716. https://doi.org/10.1128/AEM.02702-16 Google Scholar
- Chalkiadakis E, Dufourcq R, Schmitt S, Brandily C, Kervarec N, Coatanea D, Amir H, Loubersac L, Chanteau S, Guezennec J, Rouzeyrol M, Colin C (2013) Partial characterization of an exopolysaccharide secreted by a marine bacterium, Vibrio neocaledonicus sp. nov., from New Caledonia. J Appl Microbiol 114(6):1702–1712. https://doi.org/10.1111/jam.12184 Google Scholar
- Cojoc R, Merciu S, Oancea P, Pincu E, Dumitru L, Enache M (2009) Highly thermostable exopolysaccharide produced by the moderately halophilic bacterium isolated from a man-made young salt lake in Romania. Polish J Microbiol 58(4):289–294 ISSN: 1733-1331Google Scholar
- Dinić M, Pecikoza U, Djokić J, Stepanović-Petrović R, Milenković M, Stevanović M, Filipović N, Begović J, Golić N, Lukić J (2018) Exopolysaccharide produced by probiotic strain Lactobacillus paraplantarum BGCG11 reduces inflammatory hyperalgesia in rats. Front Pharmacol 9(1). https://doi.org/10.3389/fphar.2018.00001
- Du B, Yang Y, Bian Z, Xu B (2017) Characterization and anti-inflammatory potential of an exopolysaccharide from submerged mycelial culture of Schizophyllum commune. Front Pharmacol 8(252). https://doi.org/10.3389/fphar.2017.00252
- El Essawy AK, Abu Shady HM, Abu El Kher AM, Helal MM (2016) Antimicrobial, anticoagulation, fibrinolytic and prebiotic activities of exopolysaccharide produced by marine Klebsiella Sp. Egypt. J Exp Biol (Bot) 12(2):267–274. ISSN: 2090 – 0503. https://doi.org/10.5455/egyjebb.20161115114843 Google Scholar
- El-Deeb NM, Yassin AM, Al-Madboly LA, El-Hawiet A (2018) A novel purified Lactobacillus acidophilus 20079 exopolysaccharide, LA-EPS-20079, molecularly regulates both apoptotic and NF-κB inflammatory pathways in human colon cancer. Microb Cell Factories 17:29. https://doi.org/10.1186/s12934-018-0877-z Google Scholar
- El-Naggar ME, Abdelgawad AM, Salas C, Rojas OJ (2016) Curdlan in fibers as carriers of tetracycline hydrochloride: controlled release and antibacterial activity. Carbohydr Polym 154(10):194–203. https://doi.org/10.1016/j.carbpol.2016.08.042
- Frank JA, Reich CI, Sharma S, Weisbaum JS, Wilson BA, Olsen GJ (2008) Critical evaluation of two primers commonly used for amplification of bacterial 16S rRNA genes. Appl Environ Microbiol 74(8):2461–2470Google Scholar
- Goy RC, Britto D, Assis OBG (2009) A review of the antimicrobial activity of chitosan. Polímeros 19(3):241–247 ISSN 1678-5169Google Scholar
- Guinebretière MH, Auger S, Galleron N, Contzen M, Sarrau B, De Buyser ML, Lamberet G, Fagerlund A, Granum PE, Lereclus D, De Vos P, Nguyen-The C, Sorokin A (2013) Bacillus cytotoxicus sp. nov. is a novel thermotolerant species of the Bacillus cereus group occasionally associated with food poisoning. I J Syst Evol Microbiol 63:31–40. https://doi.org/10.1099/ijs.0.030627-0 Google Scholar
- Holt JG, Krieg NR, Sneath PHA, Staley JT, Williams ST (1994) Bergey’s manual® of determinative bacteriology, 9th edn. Williams & Wilkins, MarylandGoogle Scholar
- Kielak AM, Castellane TCL, Campanharo JC, Colnago LA, Costa OYA, Corradi da Silva ML, Veen JA, Lemos EGM, Kuramae EE (2017) Characterisation of novel Acidobacteria exopolysaccharides with potential industrial and ecological applications. Sci Rep 7(41193). https://doi.org/10.1038/srep41193
- Lowry OH, Rosbrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193(1):265–275Google Scholar
- Manca MC, Lama L, Improta R, Esposito E, Gambacorta A, Nicolaus B (1996) Chemical composition of two exopolysaccharides from Bacillus thermoantarcticus. Appl Environ Microbiol 62(9):3265–3269Google Scholar
- Moscovici M (2015) Present and future medical applications of microbial exopolysaccharides. Front Microbiol 6(1012). https://doi.org/10.3389/fmicb.2015.01012
- Okinaka RT, Keim P (2016) The phylogeny of Bacillus cereus sensu lato. Microbiol Spectr 4(1). https://doi.org/10.1128/microbiolspec.TBS-0012-2012
- Pawar ST, Bhosale AA, Gawade TB, Nale TR (2013) Isolation, screening and optimization of exopolysaccharide producing bacterium from saline soil. J Microbiol Biotechnol Res 3(3):24–31 ISSN 2231-3168Google Scholar
- Poli A, Anzelmo G, Fiorentino G, Nicolaus B, Tommonaro G, Di Donato P (2011) Polysaccharides from wastes of vegetable industrial processing: new opportunities for their eco-friendly re-use. In: Elnashar M (ed) Biotechnology of Polymers. InTech, London, pp 33–56Google Scholar
- Prochnow A, Clauson M, Hong J, Murphy AB (2016) Gram positive and gram negative bacteria differ in their sensitivity to cold plasma. Sci Rep 6(38610). doi:10.1038/srep38610Google Scholar
- Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4(4):406–425. https://doi.org/10.1093/oxfordjournals.molbev.a040454 Google Scholar
- Sardari RRR, Kulcinskaja E, Ron EYC, Björnsdóttirc S, Friðjónsson OH, Hreggviðsson GO, Karlsson EN (2017) Evaluation of the production of exopolysaccharides by two strains of the thermophilic bacterium Rhodothermus marinus. Carbohydrate Pol 156:1–8. https://doi.org/10.1016/j.carbpol.2016.08.062 Google Scholar
- Sardi JCO, Polaquini CR, Freires IA, Galvão LCC, Lazarini JG, Torrezan GS, Regasini LO, Rosalen PL (2017) Antibacterial activity of diacetylcurcumin against Staphylococcus aureus results in decreased biofilm and cellular adhesion. J Med Microbiol 66:816–824. https://doi.org/10.1099/jmm.0.00049 Google Scholar
- Schmidt TR, Scott EJ, Dyer DW (2011) Whole-genome phylogenies of the family Bacillaceae and expansion of the sigma factor gene family in the Bacillus cereus species-group. BMC Genomics 12(430). https://doi.org/10.1186/1471-2164-12-430
- Silhavy TJ, Kahne D, Walker S (2010) The bacterial cell envelope. Cold Spring Harb Perspect Biol 2(5). https://doi.org/10.1101/cshperspect.a000414
- Singha TK (2012) Microbial extracellular polymeric substances: production, isolation and applications. IOSR J Pharm 2(2):276–281. http://www.iosrphr.org/papers/v2i2/ZC022276281.pdf. Accessed 16 December 2018. https://doi.org/10.9790/3013-0220276281
- Smibert RM, Krieg NR (1994) Phenotypic characterization. In: Gerhardt P, Murray R, Wood W, Krieg N (eds) Methods for general and molecular bacteriology. ASM Press, Washington DC, pp 607–654Google Scholar
- Soria-Mercado IE, Villareal-Gomez LJ, Rivas GG, Sanchez NEA (2012) Bioactive compounds from bacteria associated to marine algae. In: Sammour RH (ed) Biotechnology - molecular studies and novel applications for improved quality of human life. In-Tech, London, pp 25–44Google Scholar
- Swofford DL (2002) PAUP*. Phylogenetic analysis using parsimony (*and other methods). Version 4, Sinauer Associates, Sunderland MassachusettsGoogle Scholar
- Trinetta V, Cutter CN (2016) Pullulan: a suitable biopolymer for antimicrobial food packaging applications. In: Barros-Velázquez J (ed) Antimicrobial food packaging. Academic Press, Waltham MA, pp 385–397Google Scholar
- Vuyst L, Vanderveken F, Van de Ven S, Degeest V (1998) Production by and isolation of exopolysaccharides from Streptococcus thermophilus grown in a milk medium and evidence for their growth-associated biosynthesis. J Appl Microbiol 84:1059–1068. https://doi.org/10.1046/j.1365-2672.1998.00445.x Google Scholar
- Wilson K (2003) Preparation of genomic DNA from bacteria. In: Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (eds) Current protocols in molecular biology. John Wiley & Sons Inc., Somerset NJ, pp 2.4.1–2.4.5Google Scholar
- Woo PCY, Teng JLL, Wu JKL, Leung FPS, Tse H, Fung AMY, Lau SKP, Yuen K (2009) Guidelines for interpretation of 16S rRNA gene sequence-based results for identification of medically important aerobic gram-positive bacteria. J Med Microbiol 58(Pt 8):1030–1036. https://doi.org/10.1099/jmm.0.008615-0 Google Scholar
- Yadav V, Prappulla SG, Jha A, Poonia A (2011) A novel exopolysaccharide from probiotic Lactobacillus fermentum CFR 2195: production, purification and characterization. Biotechnol Bioinf Bioeng 1(4):415–421 ISSN 2249-9075Google Scholar
- Yoon SH, Ha SM, Kwon S, Lim J, Kim Y, Seo H, Chun J (2017) Introducing EzBioCloud: a taxonomically united database of 16S rRNA and whole genome assemblies. Int J Syst Evol Microbiol 67:1613–1617Google Scholar