Food research and molecular biotechnology at FCUB Faculty of Chemistry, University of Belgrade, Serbia FCUB Founded in 1988 by the Government of Serbia, following division of the Faculty of Natural Sciences into several independent faculties Non-profit entity Education of chemists, biochemists and chemistry teachers; research 92 employed researchers, 42 teachers From 2009 it has its own Innovation Center with 25 employed researchers Resources National Projects (MNTR) National Investment fund Own resources (international students) International Projects Research projects: NATO SFP Mobility schemes: COST 928, DAAD Education support: TEMPUS, WUS Austria Facilities Starting from 2005 trough various funding resources, up to 1 mEuro was put into upgrading research facilities at the FCUB Protein analysis (PA) Structural Instrumental Analysis (SIA) Molecular Biology and Biotechnology (MBB) Cell Culture facilities (CC) National Investment Fund and WUS Austria grants 2003 Protein analysis 2004 Laboratory for Molecular Biology 2007 Structural instrumental analysis Cell culture laboratory 2009 Laboratory for Molecular Biotechnology Facilities for protein isolation, purification and analysis Ultracentrifuge Sorwall 80, ÄKTA purifier HPLC, 1D and 2D PAGE protein analysis, semy-dry and wet electroblotters, ELISA microplate reader and washer, Waters Breeze HPLC with electrochemical detection, Gel Documentation System, Eppendorf Centrifuges microcentrifuges with cooling, System for Ultra pure water, Deep Freezers (-80 oC), Lyophylizer. Molecular Biotechnology and Enzymology facilities Master Cyclers with gradient system, Electroporators, Autoclave, Incubators and shakers, Illuminators, Microplate readers, Centrifuges with cooling, Low pressure LC system, Deep Freezers (-80 oC), Analytical and Preparative Electrophoresis systems for DNA analysis, inNO Nitric Oxide measuring System, Innovative Instruments Inc (USA), inO2 Dissolved Oxygen meter, Innovative Instruments Inc (USA). Cell culture facilities Laminar flows, CO2 incubators, Centrifuges with cooling, Microscopes and inverted microscopes, Deep Freezers (-80 oC), Liquid nitrogen containers, Flow cytometer FACSCalibur (BD Biosciences). Structural instrumental analysis facilities Superconducting Fourier-Transform NMR Spectrometer AVANCE 500 MHz, Parr Hydrogenization system up to 200 barr, System for synthesis and purification of small physiologically active molecules consisting of glove box, microwave reactor and Biotage chromatography system, GC-MS, Elemental analysis system, UV/VIS spectrophotometer, FT-IR Spectrometer, Polarimeter, HPLC with UV/VIS detection, HPLC with electrochemical detection. Strategic plan of the FCUB development for the 5year period involves following: - Food safety and quality: Evaluation and standardization of food allergenic extracts (peanuts, cow’s milk, fruit and vegetables) at the molecular level. Development of cutting-edge methods for trace allergen contaminants determination in food matrix (MS-based and multiplex technologies based on flow-cytometry). - Food biotechnology and enzymology: Development of new enzyme tools for food industry, based on production, preparation and stabilization of enzymes (protease, glycohydrolases, lipases) by techniques of molecular biotechnology. Development of new recombinant allergens for the purposes of allergy diagnosis improvement. Translation of research to industry and SME. - Food for health: Development of new prebiotic and probiotic products for healthy newborns - In vitro infant formulae. New nutraceuticals development with organically bounded trace elements. Healthy diet: methods development for organically bounded oligo- and micro-elements in food. Impact of bioprocessing of food on health effects of food, digestion, allergenicity and bioactive compounds release.Translation of research to industry and SME. Research groups presentation Department of Biochemistry Department of Analytical Chemistry Research group 1 (RG1): Main area of interest: Group leader: Tanja Cirkovic Velickovic PhD students: Jelena Radosavljevic, Luka Mihajlovic, Jana Ognjenovic, Marija Stojadinovic MS students: Jasna Bojcevski ● Food bioprocessing ● Health effects of food ● Food safety, including food allergens Ongoing projects: Microwave and ultrasound enhanced action of cross-linking enzymes on proteinaceous substrates Effects of polysaccharide food matrices and bioprocessing on the protein digestion by gastrointestinal enzymes Food allergens and hypoallergenic foods by chemical and enzymatic means New quantification methods for food allergen detection, including multiplex technology Recombinant food allergens for food allergy diagnosis Research group 2 (RG2): Main area of interest: Group leader: Marija Gavrovic-Jankulovic Senior researchers: Milan Nikolic PhD students: Milica Popovic, Milica Grozdanovic, Rajna Dimitrijevic, Mohamed Salem Alzwam Abugren MS students: Igor Macinkovic, Draidi Abuajila Rogaia ● Food nutrients with immunomodulatory potential ● Food safety including food allergens ● Food probiotics ● Health effects of food Ongoing projects: Modulation of human immune response by food nutrients Production of food proteins by recombinant DNA technology Identification of novel food allergens & development of a monoclonal-antibody based assay for detection of food allergens Microbial production of novel food probiotics Influence of the different dietary patterns on red blood cell lipids Research group 3 (RG3): Main area of interest: Group leaders: Ratko Jankov PhD students: Dusan Velickovic, Aleksandra Dimitrijevic, Milena Rikalovic, Ziyad Tantoush ● Enzyme isolation and characterization ● Enzyme stabilization and application in nonaqueos systems Ongoing projects: Isolation, immobilization and stabilization of glucosidases (glucoamylases, α-glucosidases, lipases) Synthesis of physiologycally active glycosides Application of glycosidases and lipases in synthesys of glycolipids and glycosides. Food bioprocessing by oxidases Research group 4 (RG4): Main area of interest: Group leader: Zoran Vujcic PhD students: Uros Andjelkovic, Biljana Dojcinov, Aleksandra Milovanovic, Nikola Loncar ● Food bioprocessing ● Food safety, including food contaminants ● Food utilization ● Animal feedstuffs Ongoing projects: Enzyme immobilization techniques (using natural, synthetic and composed materials) and their use in continual substrate biotransformations (bioreactors) and biosensors. New quantification methods for food enzyme and contaminant detection. Chemical modifications of proteins and nucleic acids; examination of effects of modification on the structure and function of proteins and nucleic acids. Food chemistry and biochemistry. Flavours. Connection between chemistry of natural products with food chemistry and natural food contaminants. Purification and characterization of enzymes from various food sources. Examination of enzyme structures and their interactions with other proteins (antibodies, lectins, enzymes and enzyme inhibitors) and nucleic acids. Influence of food ingredients on the digestive enzyme expression and food utilization with midgut of Morimus funereus larvae as a model system. Research group 5 (RG5): Main area of interest: Group leader: Miroslav Vrvic Senior researchers: Ljuba Mandic, Vesna Jovanovic PhD students: Mila Ilic, Nikoleta Lugonja, Olga Martinov, Vladimir Beskoski, Srdjan Miletic, Jelena Milic, Jelena Acimovic,Tamara Lazarevic-Pasti ● Food biotechnology ● Food supplements ● Food microbiology ● Food safety control ● Health effects of food Ongoing projects: Developing of new prebiotic and probiotic products for healthy newborns - In vitro examination of infant formulae. New nutraceuticals – organically bounded trace elements: chromium yeast Examination of organically bounded oligo- and micro-elements in food – healthy diet Characterization of advanced glycation end products. Protein thiol group modification with methylglyoxal generated in diabetes. Enzymes and modified proteins – diagnostic markers of diabetic complications and chronic renal failure. Development of methods for their determinations. Research group 6 (RG6): Main area of interest: Group leader: Dusana Milojkovic-Opsenica Senior researchers: Zivoslav Tesic, Maja Natic, Dragan Manojlovic, Jelena Mutic PhD students: Jelena Trifkovic, Filip Andric, Kristina Lazarevic, Dragana Dabic, Ivan Andjelkovic Determination of trace elements in food by ICP and AAS. Trace analysis of pesticides residues by HPLC-UV detection. Chromatographic analysis (HPTLC, HPLC, IC) of food additives. Water quality control (analysis of anions) by ion chromatography. Application of bioassays for screening purposes of POPs chemicals in food. Ongoing projects: New method development and method assessment for quantification of trace elements and multiresidue analysis of pesticides in foodstuff. Food origin and authenticity, development and application of novel chemometrics approach. Macroporous ultramicroelectrode arrays for environmental analysis of metal cations ultratraces. Use of ELISA tests based on formation of ArH receptors–dioxin like compounds complex in screening for POPs chemicals in foodstuff. Food biotechnology and enzymology RG3 RG4 RG5 RG1 RG6 Food quality and safety RG2 Health effects of food Selected publications: Health effects of food Gavrovic-Jankulovic, M., Grozdanovic, M. (in press 2010) Application of Plant-Derived Food Lectins in Proteoglycomics and Immunomodulation. Handbook of Nutritional Biochemistry: Genomics, Metabolomics and Food Supply. Ed.: Sondre Haugen and Simen Meijer ISBN: 978-1-60741-916-7 Nikolic, M., Medic Ristic, D., Stanic, D., Postic, M., Arsic, A., Niketic, V. (2008). Dietary lipid intake influences the level of cholesterol bound to haemoglobin in human erythrocytes. European Journal of Nutrition. 47(3), 123-130. (RG2) Gorjanovic S., Beljanski M., Gavrovic-Jankulovic, M., Gojgic-Cvijovic G., Pavlovic M., Bejosano F. (2007) Antimicrobial activity of malting barley grain thaumatin-like protein isoforms S and R, Journal of the Institute of Brewing, 113, 206-212. (RG5) Gavrovic-Jankulovic, M., Poulsen, K., Brckalo, T., Bobic, S., Lindner, B., Petersen, A. (2008) A novel recombinantly produced banana lectin isoform is a valuable tool for glycoproteomics and a potent modulator of the proliferation response in CD3+, CD4+, and CD8+ populations of human PBMCs. International Journal of Biochemistry and Cell Biology 40(5), 929-41. (RG2) Dimitrijevic, R., Stojanovic, M., Zivkovic, I., Petersen, A., Jankov, R.M., Dimitrijevic, Lj., Gavrovic-Jankulovic, M. (2009) The identification of a low molecular mass bacteriocin rhamnosin A, produced by Lactobacillus rhamnosus strain 68. Journal of Applied Microbiology doi: 10.1111/j.1365-2672.2009.04539.x (RG2) Jovanovic, B.V., Dimitrijevic-Sreckovic, S.V., Mandic, M.Lj. (2008) Serum N-acetyl--D-glucosaminidase profiles in type 1 diabetes secondary complications: causes of changes and significance of determination. Journal of Clinical Laboratory Analysis, 22, 307-313. (RG5) Nikolic, M., Vranic, D., Spiric, A., Batas, V., Nikolic-Kokic, A., Radetic, P., Turubatovic, L., Blagojevic, D.P., Jones, D.R., Niketic, V., Spasic, M.B. (2006) Could cholesterol bound to haemoglobin be a missing link for the occasional inverse relationship between superoxide dismutase and glutathione peroxidase activities? Biochemical Biophysical Research Communication, 348(1), 265-270. (RG2) Selected publications: Food quality and safety Polovic, N., Pjanovic, R., Burazer, L., Velickovic, S., Jankov, R., Cirkovic Velickovic, T. (2009) Acid-formed pectin gel delays major incomplete kiwi fruit allergen Act c 1 proteolysis in in vitro gastrointestinal digestion, Journal of the Science of Food and Agriculture 89(1), 8-14. (RG1) Polovic, N., Blanusa, M., Gavrovic-Jankulovic, M., Atanaskovic-Markovic, M., Burazer, L., Jankov, R., Cirkovic Velickovic, T. (2007) A matrix effect in pectin-rich fruits hampers digestion of allergen by pepsin in vivo and in vitro. Clinical and Experimental Allergy, 37(5), 764-71. (RG1) Blanusa, M., Perovic, I., Popovic, M., Polovic, N., Burazer, L., Milovanovic, M., Gavrovic-Jankulovic, M., Jankov, R., Cirkovic Velickovic, T. (2007) Quantification of Art v 1 and Act c 1 being major allergens of mugwort pollen and kiwi fruit extracts in mass-units by ion-exchange HPLC-UV method. Journal of Chromatography, B: Analytical Technologies in the Biomedical and Life Sciences, 857(2), 188-194. (RG1) Gavrovic-Jankulovic, M., Spasic, M., Cirkovic Velickovic, T., Stojanovic, M., Inic-Kanada, A., Dimitrijevic, L., Lindner, B., Petersen, A., Becker, W.M., Jankov, R.M. (2008) Quantification of the thaumatin-like kiwi allergen by a monoclonal antibody-based ELISA. Molecular Nutrition Food Research, 52(6), 701-7. (RG2) Popovic, M., Milovanovic, M., Burazar, L., Vuckovic, O., Hoffmann-Sommergruber, K., Knulst, A., Lindner, B., Petersen, A., Jankov, R., Gavrovic-Jankulovic, M. (accepted May 2009) Cysteine proteinase inhibitor Act d 4 is a functional allergen contributing to the clinical symptoms of kiwifruit allergy. Molecular Nutrition Food Research. (RG2) Vidovic, S., Stojanovic, B., Veljkovic, J., Prazic-Arsic, L., Roglic, G., Manojlovic, D. (2008) Simultaneous determination of some water-soluble vitamins and preservatives in multivitamin syrup by validated stability-indicating high-performance liquid chromatography method. Journal of Chromatography A, 1202(2), 155-162. (RG6) Stanic D., Monigioudi E. Dilek E., Radosavljevic J., Lantto R., Mattinnen M., Vuckovic O., Bucher J., Cirkovic Velickovic T (2009) Allergenicity and digestibility of cross-linked beta-casein. Molecular Nutrition Food Research, in press. (RG1) Selected publications: Molecular Biotechnology and Enzymology Milosavic, N., Prodanovic, R., Jankov, R.M. (2007) A simple and efficient one-step, regioselective, enzymatic glucosylation of arbutin by a-glucosidase. Tetrahedron Letters, 48, 7222-7224. (RG3) Milosavic, N., Prodanovic, R., Jovanovic, S., Vujcic, Z. (2007) Immobilization of glucoamylase via its carbohydrate moiety on macroporous poly(GMA-co-EGDMA). Enzyme and Microbial Technology, 40, 14221426. (RG3) Milovanovic, A., Bozic, N., Vujcic, Z. (2007) Cell wall invertase immobilization within calcium alginate beads. Food Chemistry, 104(1), 81-86. (RG4) Vujcic, Z., Dojnov, B., Milovanovic, A., Bozic, N. (2008) Purification and Properties of the Major Leucyl Aminopeptidase from Solanum tuberosum Tubers. Fruit, Vegetable and Cereal Science and Biotechnology, 2(1), 125-130. (RG4) Dojnov, B., Bozic, N., Nenadovic, V., Ivanovic, J., Vujcic, Z. (2008) Purification and properties of midgut αamylase isolated from Morimus funereus (Coleoptera: Cerambycidae) larvae. Comparative Biochemistry and Physiology, 149B, 153-160. (RG4) Bozic, N., Ivanovic, J., Nenadovic, V., Bergstrom, J., Larsson, T., Vujcic, Z. (2008) Purification and properties of major midgut leucyl aminopeptidase of Morimus funereus (Coleoptera, Cerambycidae) larvae. Comparative Biochemistry and Physiology 149B, 454–462. (RG4) Momic, T., Vujcic, Z., Vasic, V. (2008) Kinetics of Inhibition of Peroxidase Activity of Myeloperoxidase by Quercetin. International Journal of Chemical Kinetics, 40(7), 384-394. (RG4) Radulovic, M., Cvetkovic, O., Nikolic, S., Djordjevic, D., Jakovljevic, D., Vrvic, M. (2008) Simultaneous production of pullulan and biosorption of metals by Aureobasidium pullulans strain CH-1 on peat hydrolisate. Bioresource Technology, 99, 6673-6677. (RG5) Ilic, S., Konstatinovic, S., Veljkovic, V., Savic, D., Lazic, M., Gojgic-Cvijovic, G. (2008) Impact of carboxymethylcellulose on morphology and antibiotic production by Streptomyces hygroscopicus. Current Microbiology, 57, 8-11. (RG5) Stanic D., Radosavljevic J., Polovic N., Kostic-Jadranin M., Cirkovic Velickovic T., Removal of N-terminal peptide from beta-lactoglobulin by contaminants in phenol oxidase preparations. (2009) International Dairy Journal, in press (RG1)