52 Journal on Processing and Energy in Agriculture 19 (2015) 1 Biblid: 1821-4487 (2015) 19; 1; p 52-55 Original Scientific Paper UDK: 633.21 Originalni naučni rad WHITE WINE PRODUCTION EFFLUENTS USED FOR BIOTECHNOLOGICAL PRODUCTION OF XANTHAN EFLUENTI IZ PROIZVODNJE BELOG VINA U BIOTEHNOLOŠKOJ PROIZVODNJI KSANTANA Bojana BAJIĆ, Zorana RONČEVIĆ, Vladimir PUŠKAŠ, Uroš MILJIĆ, Siniša DODIĆ, Jovana GRAHOVAC, Jelena DODIĆ University of Novi Sad, Faculty of Technology, Department of Biotechnology and Pharmaceutical Engineering, Bulevar cara Lazara 1, 21000 Novi Sad, Serbia e-mail: baj@uns.ac.rs ABSTRACT This paper examines xanthan production by Xanthomonas campestris on wastewater obtained from different white wine production process sections – crushing, pressing, clarification of must and fermentation. Wastewater was analyzed for basic nutrients content and, if required it was enriched or diluted to contain amounts suitable for xanthan biosynthesis. Biosynthesis was performed in a laboratory-scale bioreactor under identical conditions for all four studied wastewaters. In order to determine if the performed biosynthesis was successful, xanthan yield was determined gravimetrically and sugar conversion was calculated based on initial and residual HPLC values of sugar content. Obtained xanthan yield was in the range of 4.0 to 10.67 g/L while the value of sugar conversion ranged between 62.74 and 68.83 %. Different viscosities of the cultivation media at the end of the process were also compared. Due to the fact that the synthesized xanthan had both different yields and viscosities, it can be concluded that different fractions of winery wastewater yield different amounts of xanthan of different quality. Key words: environmental protection, winery wastewaters, biotechnological process, xanthan production, Xanthomonas campestris. REZIME Otpadne vode prehrambene i industrije pića, među kojima se nalaze i značajne količine otpadnih voda dobijenih proizvodnjom vina, predstavljaju nezanemarljiv ekološki problem. Najveće količine otpadnih voda vinarija se generišu u jesenjem periodu, od kraja avgusta do novembra, kada traje berba i prerada grožđa. U cilju zaštite životne sredine i obrade značajnih količina generisanog otpada neophodno je istraživati i razvijati nove procese među kojima značajno mesto zauzimaju biotehnološki procesi. Biotehnološka proizvodnja ksantana na otpadnim vodama vinarije neće samo dovesti do smanjenja organskog i neorganskog opterećenja životne sredine već će takođe rezultovati sintezom novog visokovrednog proizvoda. U okviru ovog rada je ispitana proizvodnja ksantana primenom Xanthomonas campestris na četiri frakcije otpadne vode dobijene nakon različitih faza proizvodnje belog vina – muljanja, presovanja, bistrenja šire i fermentacije. Otpadne vode su analizirane u pogledu sadržaja osnovnih nutrijenata i ukoliko je potrebno, obogaćene su ili razblažene do sadržaja koji je pogodan za biosintezu ksantana. Biosinteza je izvedena u laboratorijskom bioreaktoru pod identičnim uslovima za sve četiri frakcije otpadne vode. Kako bi se utvrdilo da li je izvedena biosinteza bila uspešna određen je prinos ksantana i stepen konverzije šećera. Dobijeni prinosi su se nalazili u opsegu između 4,00 i 10,67 g/l dok je vrednost stepena konverzije iznosila od 62,74 do 68,83%. Pored toga, upoređene su i vrednosti viskoziteta kultivacionih tečnosti na kraju procesa proizvodnje ksantana. Usled činjenice da je sintetisan ksantan imao različit prinos i različit viskozitet može se zaključiti da se na različitim frakcijama otpadnih voda vinarije dobija različita količina ksantana različitog kvaliteta. Ključne reči: zaštita životne sredine, otpadne vode vinarije, biotehnološki proces, proizvodnja ksantana, Xanthomonas campestris. INTRODUCTION In order to prevent environmental pollution and process significant amounts of generated wastewaters which are usually difficult and expensive, it is necessary to research and develop new processes (Buyukgungor and Gurel, 2009). As opposed to chemical and physical methods, biological wastewater treatments can more effectively cope with a broad range of effluents and is especially suitable for the treatment of wastewater that contains common organic pollutants, which is why biotechnology is the most commonly used technology for wastewater treatment. Biotechnological utilization of wastes and wastewaters can be used for the production of products of additional value and minimization of waste (Mojović et al., 2014; Pejin et al., 2014). Food and beverage industries, including the winemaking industry generate significant amounts of wastewaters that are a large environmental pollutant (Klašnja and Šćiban, 2000). The amount of wastewater varies between different wineries and different years, but is in the range of 0.1 to 2.4 m 3 per m 3 of produced wine (Brito et al., 2007). Wineries produce large amounts of wastewater resulting from numerous cleaning operations that occur during the production stages of wine. This wastewater originates from washing operations after the crushing and pressing of grapes, as well as rinsing of fermentation tanks, barrels and other equipment or surfaces. Therefore, wastewaters from different fractions will necessarily have different characteristics and components, and the most predominant compounds in these kinds of wastewaters are soluble sugars (fructose and glucose), organic acids (tartaric, lactic and acetic), alcohols (glycerol and ethanol) and high molecular weight compounds, such as polyphenols, tannins and lignin (Souza et al., 2013; Chapman et al., 2001). The resulting effluent contains various organic and inorganic substances and its environmental impact is significant, mainly due to its high organic and inorganic load, the large volumes produced and its seasonal variability (Petruccioli et al., 2000; Ioannou et al., 2015). An extracellular microbial polysaccharide, xanthan is produced by bacteria of the genus Xanthomonas. Due to its rheological properties, xanthan has a wide range of applications in food and non-food industries as a stabilizer, thickener, emulsifier and suspending agent. Xanthan gum produces highly