Open Access

Peer-reviewed

Research Article

Main Article Content

Subba Rao Toleticorresponding author
Senthil Kumar
Sharat Chandra

Abstract

Two Pseudomonas aeruginosa isolates from natural biofilms (marine and freshwater environment) were investigated for exopolymer (EPS) production and chemical characterization.  Both the isolates were categorized using molecular tools as P. aeruginosa species, The EPS production was distinct with various carbon and nitrogen sources, the average EPS yield by the two Pseudomonas strains was 55 µg ml-1.  Modelling was done to establish the relation between observed and predicted EPS yields. The chemical composition, FTIR and Raman spectroscopy analysis of the two EPS showed that carbohydrate content was more in marine strain, while protein content was relatively high in the freshwater strain. Thermo-gravimetric analysis of the two EPS showed endothermic decomposition. Biochemical study by gel permeation chromatography showed that the marine strain EPS is a highly glycosylated biomolecule, while the freshwater EPS is a weakly glycosylated biomolecule with molecular weights 140,000 and 300,000 Daltons respectively. The EPS produced by the two Pseudomonas isolates has implication in process and chemical industries.   

Keywords
bacteria, EPS, carbohydrate, protein, biofilm, biofouling, industries

Article Details

How to Cite
Toleti, S. R., Kumar, S., & Chandra, S. (2020). Comparative assessment of exopolymer production and chemical characteristics of two environmental biofilm isolates of Pseudomonas aeruginosa. Chemical Reports, 2(1), 132-143. https://doi.org/10.25082/CR.2020.01.003

References

  1. Flemming HC andWingender J. The biofilm matrix. Nature Reviews Microbiology, 2010, 8(9): 623-633. https://doi.org/10.1038/nrmicro2415
  2. Simon M, Grossart HP, Schweitzer B, et al. Microbial ecology of organic aggregates in aquatic ecosystems. Aquatic Microbial Ecology, 2002, 28: 75-211. https://doi.org/10.3354/ame028175
  3. Nwodo U, Green E and Okoh A. Bacterial exopolysaccharides: functionality and prospects. International Journal of Molecular Sciences, 2012, 13(11): 14002-14015. https://doi.org/10.3390/ijms131114002
  4. Ha YW, Stack RJ, Hespell RB, et al. Some chemical and physical properties of extracellular polysaccharides produced by Butyrivibrio fibrisolvens strains. Applied and Environmental Microbiology, 1991, 57(7): 2016-2020. https://doi.org/10.1128/AEM.57.7.2016-2020.1991
  5. Abu GO,Weiner RM, Rice J, et al. Properties of an extracellular adhesive polymer from the marine bacteria Shewanella colwelliana. Biofouling, 1991, 3: 69-84. https://doi.org/10.1080/08927019109378163
  6. Christensen BE. The role of extracellular polysaccharides in biofilms. Journal of Biotechnology, 1989, 10(3-4): 181-202. https://doi.org/10.1016/0168-1656(89)90064-3
  7. Rao TS. Biofouling in IndustrialWater Systems: In: Amjad Z, Demadis, KD, editors. Mineral Scales and Deposits, The Netherlands, Elsevier, 2015, 123-140. https://doi.org/10.1016/B978-0-444-63228-9.00006-1
  8. Evans LR and Linker A. Production and characterization of the slime polysaccharide of Pseudomonas aeruginosa. Journal of Bacteriology, 1973, 116(2): 915-924. https://doi.org/10.1128/JB.116.2.915-924.1973
  9. Costerton JW, Lewandowski Z, Caldwell DE, et al. Lappinscott, Microbial biofilms. Annu. Rev. Microbiology, 1995, 49: 711-745. https://doi.org/10.1146/annurev.mi.49.100195.003431
  10. Sage A, Linker A, Evans LR, et al. Hexose phosphate metabolism and exopolysaccharide formation in Pseudomonas cepacia. Current Microbiology, 1990, 20: 191-198. https://doi.org/10.1007/BF02091996
  11. Suresh Kumar A, Mody K and Jha B. Bacterial exopolysaccharides-a perception. Journal of Basic Microbiology, 2007, 47(2): 103-117. https://doi.org/10.1002/jobm.200610203
  12. Rehm BH. Bacterial polymers: biosynthesis, modifications and applications. Nature Reviews Microbiology, 2010, 8: 578-592. https://doi.org/10.1038/nrmicro2354
  13. Freitas F, Alves VD and Reis MA. Advances in bacterial exopolysaccharides: from production to biotechnological applications. Trends in Biotechnology, 2011, 29(8): 388-398. https://doi.org/10.1016/j.tibtech.2011.03.008
  14. Nielsen PH, Jahn A and Palmgren R. Conceptual model for production and composition of exopolymers in biofilms. Water Science and Technology,1997, 36(1): 11-19. https://doi.org/10.2166/wst.1997.0002
  15. Flemming HC. EPS-then and now. Microorganisms, 2016, 4(4): 41-47. https://doi.org/10.3390/microorganisms4040041
  16. Weisburg WG, Barns SM, Pelletier DA, et al. 16S ribosomal DNA amplification for phylogenetic study. Journal of Bacteriology, 1991, 173(2): 697-703. https://doi.org/10.1128/JB.173.2.697-703.1991
  17. Tamura K, Peterson D, Peterson N, et al. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology and Evolution, 2011, 28(10): 2731-2739. https://doi.org/10.1093/molbev/msr121
  18. Jiao Y, Cody GD, Harding AK, et al. Characterization of extracellular polymeric substances from acidophilic microbial biofilms. Applied and Environmental Microbiology, 2010, 76(9): 2916-2922. https://doi.org/10.1128/AEM.02289-09
  19. Cui JD and Zhang B. Comparison of culture methods on exopolysaccharide production in the submerged culture of Cordyceps militaris and process optimization. Letters in Applied Microbiology, 2011, 52(2): 123-128. https://doi.org/10.1111/j.1472-765X.2010.02987.x
  20. Lowry OH, Rosebrough NJ, Farr AL, et al. Protein measurement with the Folin phenol reagent. The Journal of Biological Chemistry, 1951, 193(1): 265-275.
  21. Dubois M, Gilles KA, Hamilton JK, et al. Colorimetric method for determination of sugars and related substances. Analytical Chemistry, 1956, 28(3): 350-356. https://doi.org/10.1021/ac60111a017
  22. Dische Z. A new specific color reaction of hexuronic acids. The Journal of Biological Chemistry, 1947, 167: 189-198.
  23. Muralidharan J and Jayachandran S. Physicochemical analyses of the exopolysaccharides produced by a marine biofouling bacterium, Vibrio alginolyticus. Process Biochemistry, 2003, 38: 841-847. https://doi.org/10.1016/S0032-9592(02)00021-3
  24. Lee WY, Park Y, Ahn JK, et al. Factors influencing the roduction of endopolysaccharide and exopolysaccharide from Ganoderma applanatum. Enzyme and Microbial Technology, 2007, 40(2): 249-254. https://doi.org/10.1016/j.enzmictec.2006.04.009
  25. Poli A, Anzelmo G and Nicolaus B. Bacterial exopolysaccharides from extreme marine habitats: production, characterization and biological activities. Marine Drugs, 2010, 8(6): 1779-1802. https://doi.org/10.3390/md8061779
  26. Mangwani N, Shukla SK, Rao TS, et al. Calcium-mediated modulation of Pseudomonas mendocina NR802 biofilm influences the phenanthrene degradation. Colloids and Surfaces B: Biointerfaces. 2014, 114: 301-309. https://doi.org/10.1016/j.colsurfb.2013.10.003
  27. Shukla SK and Rao TS. Effect of calcium on Staphylococcus aureus biofilm architecture: a confocal laser scanning microscopic study. Colloids and Surfaces B: Biointerfaces. 2013, 103: 448-454. https://doi.org/10.1016/j.colsurfb.2012.11.003
  28. Elisashvili VI, Kachlishvili ET and Wasser SP. Carbon and nitrogen source effects on basidiomycetes exopolysaccharide production. Applied Biochemistry and Microbiology, 2009, 45(5): 531-535. https://doi.org/10.1134/S0003683809050135
  29. Nichols PD, Henson JM, Guckert JB, et al. Fourier transform infrared spectroscopic methods for microbial ecology: analysis of bacteria, bacteri-polymer mixtures and biofilms. Journal of Microbiological Methods, 1985, 4: 79-94. https://doi.org/10.1016/0167-7012(85)90023-5
  30. Nyquist RA. Interpreting infrared, Raman, and nuclear magnetic resonance spectra. Academic Press. New York, 2001.
  31. Hsieh C, Tsai MJ, Hsu TH, et al. Medium optimization for polysaccharide production of Cordyceps sinensis. Applied Biochemistry and Biotechnology, 2005, 120(2): 145-157. https://doi.org/10.1385/ABAB:120:2:145
  32. Dertli E, Mayer MJ and Narbad A. Impact of the exopolysaccharide layer on biofilms adhesion and resistance to stress in Lactobacillus johnsonii FI9785. BMC Microbiology, 2015, 15: 8. https://doi.org/10.1186/s12866-015-0347-2
  33. Laspidou CS and Rittmann BE. Modeling the development of biofilm density including active bacteria, inert biomass, and extracellular polymeric substances. Water Research, 2004, 38(14-15): 3349-3361. https://doi.org/10.1016/j.watres.2004.04.037
  34. Lion LW, Shuler ML, Hsieh KM, et al. Trace metal interactions with microbial biofilms in natural and engineered systems. Critical Reviews in Environmental Science and Technology, 1988, 17(4): 273-306. https://doi.org/10.1080/10643388809388338
  35. Nwodo UU, Green E and Okoh AI. Bacterial Exopolysaccharides: Functionality and Prospects. International Journal of Molecular Sciences, 2012, 13, 14002-14015. https://doi.org/10.3390/ijms131114002
  36. Moscovici M. Present and future medical applications of microbial exopolysaccharides. Frontiers in Microbiology, 2015, 6(1012): 1-11. https://doi.org/10.3389/fmicb.2015.01012