A statistical approach on optimization of exopolymeric substance production by Halomonas sp. S19 and its emulsification activity

Abstract

An exopolymer producing bacterial strain was identified as Halomonas sp. S19 by 16S rRNA gene sequencing isolated from Mandapam, Southeast coast of India. Strain S19 produces a significant amount of exopolymer (320 mg L⁻¹) in a medium optimized with 2.5 % glucose, 0.6 % peptone, 7.5 % salt and pH 7.5 at 35 ^°C. The exopolymer consists of total sugars (65 %), proteins (4.07 %), uronic acids (8.08 %) and sulphur contents (6.39 %). FT-IR and ¹H NMR analysis revealed the presence of functional groups corresponding to carbohydrates, proteins and sulphates. The exopolymer of Halomonas sp. S19 emulsifies different oils. However, 10 % exopolymer shows 55.18, 55.18, 49.81 and 24.62 % of emulsifying activity for sesame oil, coconut oil, paraffin and kerosene. The present study was focused on optimisation of exopolymer production using Box–Behnken experimental design and its possibility for potential emulsification index.

Keywords

Halomonas sp. / Exopolymer / Optimisation / NMR / DSC / Emulsification

References

Abou-El-Hossein KA, Kadirgamaa K, Hamdib M, Benyounis KY. Prediction of cutting force in end-milling operation of modified AISI P20 tool steel. J Mater Process Tech, 2007, 182: 241-247.

Arias S, Del Moral A, Ferrer MR, Tallon R, Quesada E, Bejar V. Mauran, an exopolysaccharide produced by the halophilic bacterium Halomonas maura, with a novel composition and interesting properties for biotechnology. Extremophiles, 2003, 7: 319-326.

Bach H, Gutnick DL. Novel polysaccharide-protein based amphipathic formulations. Appl Microbiol Biotechnol, 2005, 71: 34-38.

Banat IM, Makkar RS, Cameotra SS. Potential commercial applications of microbial surfactants. Appl Microbiol Biotechnol, 2000, 53: 495-508.

Bouchotroch S, Quesada E, Izquierdo I, Rodriguez M, Bejar V. Bacterial exopolysaccharides produced by newly discovered bacteria belonging to the genus Halomonas, isolated from hypersaline habitats in Morocco. J Ind Microbiol Biotechnol, 2000, 24: 374-378.

Bramhachari PV, Kishor PB, Ramadevi R, Kumar R, Rao BR, Dubey SK. Isolation and characterization of mucous exopolysaccharide (EPS) produced by Vibrio furnissii strain VBOS3. J Microbiol Biotechnol, 2007, 17: 44-51.

Cooper DG, Goldenberg BG. Surface-active agents from two Bacillus species. Appl Environ Microbiol, 1987, 53: 224-229.

Decho AW. Barnes M. Microbial exopolymer secretions in ocean environments: their role(s) in food webs and marine processes. Oceanography and Marine Biology: an Annual Review, 1990, Aberdeen: Aberdeen Univ Press, 73-153.

Dodgson KS. Determination of inorganic sulphate in studies on the enzymic and nonenzymic hydrolysis of carbohydrate and other sulphate esters. Biochem J, 1961, 78: 312-319.

Dubois M, Gilles K, Hamilton J, Rebers P, Smith F. Colorimetric method for determination of sugars and related substances. Anal Chem, 1956, 28(3): 350-356.

Filisetti-Cozzzi TMCC, Carpaita NC. Measurement of uronic acid without interference of neutral sugars. Anal Biochem, 1991, 197: 157-162.

Freitas F, Alves VD, Pais J, Costa N, Oliveira C, Mafra L, Hilliou L, Oliveira R, Reis MA. Characterization of an extracellular polysaccharide produced by a Pseudomonas strain grown on glycerol. Bioresour Technol, 2009, 100: 859-865.

Gandhi HP, Ray RM, Patel RM. Exopolymer production by Bacillus species. Carbohydr Polym, 1997, 34: 323-327.

Gutierrez T, Mulloy B, Black K, Green DH. Glycoprotein emulsifiers from two marine Halomonas species: chemical and physical characterization. J Appl Microbiol, 2007, 103: 1716-1727.

Gutierrez T, Shimmield T, Haidon C, Black K, Green DH. Emulsifying and metal ion binding activity of a glycoprotein exopolymer produced by Pseudoalteromonas sp. strain TG12. Appl Environ Microbiol, 2008, 74: 4867-4876.

Jain RM, Mody K, Mishra A, Jha B. Isolation and structural characterization of biosurfactant produced by an alkaliphilic bacterium Cronobacter sakazakii isolated from oil contaminated wastewater. Carbohydr Polym, 2012, 87: 2320-2326.

Junkins AD, Doyle MP. Demonstration of exopolysaccharide production by enterohemorrhagic Escherichia coli. Curr Microbiol, 1992, 25(1): 9-17.

Kadirgama K, Abou-El-Hosseirr KA, Mohammad B, Habeeb H. Statistical model to determine surface roughness when milling hastelloy C-22HS. J Mech Sci Technol, 2007, 21: 1651-1655.

Kumar AS, Mody K, Jha B. Bacterial exopolysaccharides—a perception. J Bas Microbiol, 2007, 47: 103-107.

Kushner DJ, Kamekura M. Rodriguez-Valera F. Physiology of halophilic eubacteria. Halophilic Bacteria, 1988, Boca Raton: CRC Press, 109-140.

Llamas I, Amjres H, Mata JA, Quesada E, Bejar V. The potential biotechnological applications of the exopolysaccharide produced by the halophilic bacterium Halomonas almeriensis. les, 2012, 17: 7103-7120.

Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem, 1951, 193: 265-275.

Mandal SK, Singh RP, Patel V. Isolation and characterization of exopolysaccharide secreted by a toxic dinoflagellate, Amphidinium carterae Hulburt 1957 and its probable role in harmful algal blooms (HABs). Microbial Ecol, 2011, 62: 518-527.

Martinez Checa F, Toledo FL, El Mabrouki K, Quesada E, Calvo C. Characteristics of bioemulsifier V2-7 synthesised in culture media added of hydrocarbons: chemical composition, emulsifying activity and rheological properties. Biores Technol, 2007, 98: 3130-3135.

Mata JA, Bejar V, Llamas I, Arias S, Bressollier P, Tallon R, Urdaci MC, Quesada E. Exopolysaccharides produced by the recently described bacteria Halomonasventosae and Halomonas anticariensis. Res Microbiol, 2006, 157: 827-835.

Mata JA, Bejar V, Bressollier P, Tallon R, Urdaci MC, Quesada E, Llamas I. Characterization of exopolysaccharides produced by three moderately halophilic bacteria belonging to the family Alteromonadaceae. J Appl Microbiol, 2008, 105: 521-528.

Mishra A, Jha B. Isolation and characterization of extracellular polymeric substances from micro algae Dunaliella salina under salt stress. Bioresour Technol, 2009, 100: 3382-3386.

Neefs JM, van de Peer Y, Hendriks L, de Wachter R. Compilation of small ribosomal subunit RNA sequences. Acids Res, 1990, 18(suppl): 2237-2317.

Nguyena NK, Borkowskib JJ. New 3-level response surface designs constructed from incomplete block designs. J Stat Plan Inference, 2008, 138: 294-305.

Parikh A, Madamwar D. Partial characterization of extracellular polysaccharides from cyanobacteria. Bioresour Technol, 2006, 97: 1822-1827.

Parthiban K, Vignesh V, Thirumurugan R. Characterization and in vitro studies on anticancer activity of exopolymer of Bacillus thuringiensis S13. Afr J Biotechnol, 2014, 13(21): 2137-2144.

Perez Fernandez ME, Quesada E, Galvez J, Ruiz C. Effect of exopolysaccharide V2-7 isolated from Halomonas eurihalina on the proliferation in vitro of human peripheral blood lymphocytes. Immunopharmacol Immunotoxicol, 2000, 22: 131-141.

Perfumo A, Smyth TJP, Marchant R, Banat IM. Timmis KN. Production and roles of biosurfactant and bioemulsifiers in accessing hydrophobic substrates. Microbiology of hydrocarbons, oils, lipids and derived compounds, 2009, Heidelberg: Springer, 1502-1512.

Quesada E, Bejar V, Calvo C. Exopolysaccharide production by Volcaniella eurihalina. Experientia, 1993, 49: 1037-1041.

Roberson EB, Firestone MK. Relationship between desiccation and exopolysaccharide production in a soil Pseudomonas sp. Appl Environ Microbiol, 1992, 4: 1284-1291.

Ruiz-Ruiz C, Srivastava GK, Carranza D, Mata JA, Llamas I, Santamaria M, Quesada E, Molina IJ. An exopolysaccharide produced by the novel halophilic bacterium Halomonas stenophila strain B100 selectively induces apoptosis in human T leukaemia cells. Appl Microbiol Biotechnol, 2011, 89: 345-355.

Singh RP, Mahendra K, Shukla Avinash Mishra, Kumari P, Reddy CRK, Jha B. Isolation and characterization of exopolysaccharides from seaweed associated bacteria Bacillus licheniformis. Carbohydr Polym, 2011, 84: 1019-1026.

Singh RP, Shukla MK, Mishra A, Reddy CRK, Jha B. Bacterial extracellular polymeric substances and their effect on settlement of zoospore of Ulva fasciata. Colloids Surf B, 2013, 103: 223-230.

Spano A, Gugliandolo C, Lentini V, Maugeri TL, Anzelmo G, Poli A, Nicolaus B. A novel EPS-producing strain of Bacillus licheniformis isolated from a shallow vent off panarea Island (Italy). Curr Microbiol, 2013, 67: 21-29.

Sutherland IW. Microbial polysaccharides from gram negative bacteria. Int Dairy J, 2001, 11: 663-674.

Uhlinger DJ, White DC. Relationship between physiological status and formation of extracellular polysaccharide glycocalyx in Pseudomonas atlantica. Appl Environ Microbiol, 1983, 45: 64-70.

Willumsen PAE, Karlson U. Screening of bacteria, isolated from PAD-contaminated soils, for production of biosurfactant and bioemulsifiers. Biodegradation, 1997, 7: 415-423.