In this study, the main structural monomer units and configuration of their glycosidic linkages of a exopolysaccharide obtained from a strain of Bacillus licheniformis was investigated. The chemical structure of this polymer, after purification of crude material, was analyzed by chemical methods, planar chromatography, elemental analysis and FTIR

Gojgić-Cvijović, Gordana

Ilić, D.

Jakovljević, Dragica

Kekez, Branka

Stefanović, Jovana

Vrvić, Miroslav

Central Repository of the Institute of Chemistry, Technology and Metallurgy (CER)

        Pon HYSICAL11th InterFundamentPhyUnder UnivePThe ConfProfesSeptemBe CHEMISnational Coal and Appsical Chem the auspicersity of Beroceeding      erence is desor  Ivan Dr        ber 24-28lgrade, Ser  TRY 2012nference lied Aspectistry s of the lgrade s dicated to aganić , 2012  bia   s of       ISBN 978-86-82475-27-9 Volume 1 ISBN 978-86-82475-28-6 Volume II  Title: PHYSICAL CHEMISTRY 2012 (Proceedings) Editors: S. Anić and Ž. Čupić Published by: Society of Physical Chemists of Serbia, Studenski trg 12-16, 11158, Belgrade, Serbia Publisher: Society of Physical Chemists of Serbia For Publisher: S. Anić, President of Society of Physical Chemists of Serbia Printed by: “Jovan” Priting and Publishing Company; 200 Copies; Number of pages: 6+ 497; Format: B5; Printing finished in September 2012.  Text and Layout: “Jovan”  200- Coppy printing  V ORGANIZERS   PHYSICAL CHEMISTRY 2012  11th International Conference on  Fundamental and Applied Aspects of Physical Chemistry   Under the auspices of the University of Belgrade    Organized by The Society of Physical Chemists of Serbia  in co-operation with   Institute of Catalysis Bulgarian Academy of Sciences   Boreskov Institute of Catalysis of Siberian Branch of the Russian Academy of Sciences  Faculty of Physical Chemistry, University of Belgrade, Serbia  Institute of Chemistry Technology and Metallurgy,  University of Belgrade, Serbia  Vinča Institute, University of Belgrade, Serbia  Institute of General and Physical Chemistry, Serbia    Faculty of Pharmacy, University of Belgrade, Serbia    VI COMMITTEES  International Organizing Committee  Chairman: S. Anić (Serbia) Vice-chairmans: M. Gabrovska (Bulgaria) V. A. Sadykov (Russia) B. Adnadjević (Serbia) Members: Z. Cherkezova-Zheleva (Bulgaria), N. Cvjetićanin (Serbia), S. N. Blagojević (Serbia), M. Daković (Serbia), T. Grozdić (Serbia), Lj. Ignjatović (Serbia), D. Jovanović (Serbia), N. Jović-Jovičić (Serbia), M. Kuzmanović (Serbia), D. Marković (Serbia), N. Kovačević (Serbia), J. Marković-Dimitrić (Serbia), B. Milosavljević (USA), Š. Miljanić (Serbia), N. Miljević (Serbia), A. Milutinović-Nikolić (Serbia), M. Mojović (Serbia), Z. Mojović, (Serbia), N. Ostrovski (Serbia), I. Pašti (Serbia), C. Pona (Italy), B. Simonović (Serbia), D. Stanisavljev (Serbia), B. Šljukić (Serbia), N. Vukelić (Serbia), V. Vukojević (Sweden)   International Scientific Committee  Chairman: Ž. Čupić (Serbia) Vice-chairmans: V. N. Parmon (Russia) S. Rakovsky (Bulgaria) Members: A. Antić-Jovanović (Serbia), G. Bačić (Serbia), R. Cervellati (Italy), R. Compton (UK), V. Dondur (Serbia), Biljana Gaković (Serbia), V. Gaspar (Hungary), S. Gorodsky (Russia), V. Goussev (France), K. Hedrih (Serbia), Florou Heleny (Greece), M. Jeremić (Serbia), A. L. Kawczyñski (Poland), Lj. Kolar-Anić (Serbia), V. Kuntić (Serbia), S. Mentus (Serbia), S. Milonjić (Serbia), Š. Miljanić (Serbia), Lj. Morozova-Roche (Sweden), P. Misaelides (Greece) M. Perić (Serbia), M. Plavšić (Serbia), V. Sadykov (Russia), G. Schmitz (Belgium), I. Schreiber (Czech), P. Sevcik (Slovakia), Natalia Skorodumova (Sweden), Dalimil Snita (Czech), N. Stepanov (Russia), D. Todorović (Serbia), M.Trtica (Serbia), V.  Vasić (Serbia), D. Veselinović (Serbia), M. Zdujić (Serbia)   Local Executive Committee  Chairman: B. Adnadjević Vice-chairman: S. Blagojević Members: A. Abu Rabi-Stanković, P. Banković, N. Begović, J. Dostanić, A. Đerić, A. Ignjatović, A. Ivanović, A. Jović, D. Lončarević, J. Krstić, J. Maksimović, M. Milenković, S. Maćešić, V. Marković, B. Nedić, M. Petković, A.  Popović-Bjelić, N. Potkonjak, D. Ranković, M. Stević, I. Stojković, M. ŽunićVII Honorary Committee  Nikola Hajdin,  President of Serbian Academy of Sciences and Arts Branko Kovačević,   Rector of University of Belgrade Radivoje Mitrović Secretary of State for Education and Science Vera Dondur President of the Council for Scientific and Technological Development Vladimir Bumbaširević Dean of the Faculty of Medicine of University of Belgrade Paula Putanov,  Member of Serbian Academy of Sciences and Arts Momčilo Ristić,  Member of Serbian Academy of Sciences and Arts Ankica Antić Jovanović,  Honorary President of Society of Physical Chemists of Serbia                   J-09-P 541 CHARACTERISATION OF EXOPOLYSACCHARIDE PRODUCED BY BACILLUS SP. NS032   J. Stefanović1, D. Ilić2, B. Kekez2, G. Gojgić-Cvijović1, D. Jakovljević1 , M. Vrvić1,2      1IChTM - Department of Chemistry, University of Belgrade, Njegoševa 12, P.O. Box 473, 11000 Belgrade, Serbia (jovana_stefanovic@chem.bg.ac.rs) 2Faculty of Chemistry, University of Belgrade, Studentski trg 12-16, P.O. Box 51,  11158 Belgrade, Serbia Abstract In this study, the main structural monomer units and configuration of their glycosidic linkages of a exopolysaccharide obtained from a strain of Bacillus licheniformis was investigated. The chemical structure of this polymer, after purification of crude material, was analyzed by chemical methods, planar chromatography, elemental analysis and FTIR.  Introduction Microbial exopolysaccharides are the group of biopolymers whose structural chemistry is very complex. Usually some types of microorganisms can be characterized by the presence of several carbohydrate polymers. The economic significance of these compounds is related to the fact that some of these polymers is nowadays widely accepted products of biotechnology, while others are in various stages of research. The use of microbial polymers varies widely due to good mechanical properties for application as fiber, films, adhesives, rheology modifiers, hydrogels, emulsifiers, and drug delivery agents. Biopolymers with industrial application are often bacterial and fungal products, like levan, dextran, β-glucan, pullulan. In this work we reported a route to produce of purified exopolysaccharide produced by strain of Bacillus licheniformis, and determination of main structural characteristics of this polymer. Experimental  Exopolysaccharide was produced by strain of Bacillus licheniformis which was isolated from petroleum sludge sample taken from Oil Refinery Novi Sad. The strain was identified using 16S rRNA gene sequence  analysis [1]. The organism was cultivated in sucrose broth for 10 days at 28 oC [2]. After removal of biomass the crude polysaccharide was isolated from fermentation culture by precipitation with two volumes of ethanol. Polymer was analyzed for its carbon, hydrogen and nitrogen content using the Vario EL III device (GmbH Hanau Instruments, Germany). Nitrogen content usually was indication for the presence of protein as impurities. After treatments with Dnase, Rnase and Pronase and dialysis, purified polysaccharide was precipitated by acetone and lyophilized. In this way was obtained purified polysaccharide, without nitrogen, with content of C and H that corresponded to polysaccharide molecules (Table 1.). Monomer components was determined by planar chromatography of the acid hydrolizate of the glycan. The ATR-IR spectrum of pure polysaccharide was obtained J-09-P 542 using Thermo Nicolet 6700 FT-IR Spectrophotometer with Smart Orbit Diamond ATR (attenuated total reflectance) accessory.   Results and Discussion Planar chromatography of the hydrolyzed crude polysaccharide, comparing with standards of authentic monosaccharides, showed two main components, fructose and glucose, in about equal proportions. After further treatments, in hydrolysate of purified polymer was detected only one component, D-Fructose, which indicated that investigated polymer is fructan. Additional evidence that purification of polysaccharide was successful was found in the results of elemental analysis of crude and purified polymer (Table 1). The FT-IR spectrum of the purified polysaccharide showed the spectral pattern typical for polysaccharides. As shown in Fig.1, absorption bands between 1128 cm-1 and 1014 cm-1 corresponded to C-O-C and C-O-H stretching vibrations [3], the bands in the range of 1200-1500 cm-1 ascribed to C-H deformation vibration, the band at 1645 cm-1 was due to bound water. Strong, wide band at 3000–3500 cm−1 and band at 2936 cm-1 was assigned to the hydroxyl groups stretching vibration of polysaccharide and C-H stretching [4]. Absorption band in the anomeric region (950-700 cm-1) at 891 cm-1 is specific for the β-configuration of the glycosidic linkages. Based on the FT-IR spectra it was concluded that investigated polymer had β-configuration of glycosidic linkages.   Table 1.  Elemental chemical composition.  % N % C % H Crude sample 7.44 30.58 5.13 Purified sample / 44.42 6.20     Figure 1. FTIR spectrum of purified exopolysaccharide produced by B. licheniformis.  Conclusion  The present study showed, on the basis of chemical methods, planar chromatography, elemental analysis and FTIR, that investigated exopolysaccharide is homopolymer, J-09-P 543 composed of a D-fructose, as  the O-specific monomer, with β-configuration of the glycosidic linkages.  Acknowledgement. This work was supported by the Ministry of Education and Science of the Republic of Serbia through Project  III 43004.  References  [1] G. D. Gojgic-Cvijovic, J. S. Milic, T. M. Solevic, V. P. Beskoski, M. V. Ilic, L. S. Djokic, T. M. Narancic, M. M. Vrvic, Biodegradation, 2012, 23, 1-14. [2]. J. Liu, J. Luo, Y. Sun, H.Ye, Z. Lu, X. Zeng, Biores. Technol., 2010, 101, 6077-6083. [3] M. Wu, Y. Wu, J. Zhou, Y. Pan, Food Chemistry, 2009, 113,1020–1024. [4] M. Mathlouthi, J. L. Koenig, Adv. Carbohydr. Chem. Biochem., 1987, 44, 7–89. 

Characterization of exopolysaccharide produced by Bacillus sp. NS032

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Characterization of exopolysaccharide produced by Bacillus sp. NS032

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