ORIGINAL PAPER Diversity and oenological characterization of indigenous Saccharomyces cerevisiae associated with Z ˇ ilavka grapes Sandi Orlic ´ • Tanja Vojvoda • Katarina Huic ´ Babic ´ • F. No ´e Arroyo-Lo ´pez • Ana Jeromel • Bernard Kozina • Lucilla Iacumin • Giuseppe Comi Received: 18 August 2009 / Accepted: 16 January 2010 / Published online: 3 February 2010 Ó Springer Science+Business Media B.V. 2010 Abstract The aim of this research was to identify the Saccharomyces spp. associated with Z ˇ ilavka grapes and to evaluate their enzymatic activities, H 2 S production and micro-fermentation performance. For this purpose, a total of 143 yeast strains isolated from three production areas of the Mostar wine region (Bosnia and Herzegovina) were studied and analysed. Firstly, yeasts were identified to genus level by growth on WL nutrient agar and the test of assimilation of lysine. Later, molecular identification at species level was carried out with RFLP analysis of 18S rDNA ? ITS region, and at strain level with microsatellite- primed PCR (MSP-PCR). At physiological level yeast strains were grouped into different clusters by means of the Joining-Tree-Clustering-Method. All yeasts tested were identified as S. cerevisiae, resulting a total of 18 different strains. All of the investigated strains produced hydrogen sulphide, 89% were able to complete the fermentation, and none of them was able to synthesize killer toxins. Since enzymes play a very important role in wine aroma development, it was very encouraging that 33% of the strains were able to synthesize pectinolytic enzyme but only one produced b-glucosidase. In the second part of the selection process two indigenous strains were compared with commercial yeast in a microvinification and Z ˇ ilavka wines with different profiles of volatiles were obtained. This research represents a first step in the selection of indigenous yeast strains from the Mostar region with the goal of maintaining the specific organoleptic characteristics of Z ˇ ilavka wine. Keywords Saccharomyces Á MSP-PCR Á Biodiversity Á Enological characterization Á Wine Introduction Saccharomyces is a yeast genus with great importance in the complex vinification process. Its predominance during spontaneous alcoholic fermentation of grape musts con- tributes to the physical, chemical and sensorial characters of wines (Pretorius 2000). The primary role of wine yeasts is to catalyse the rapid, complete and efficient conversion of grape sugar into ethanol, carbon dioxide, glycerol and other minor, but important, metabolites without the development of off flavours (Pretorius 2000). Wine fer- mentation is conducted by a variable number of Saccha- romyces strains (Agnolucci et al. 2007; Martı ´nez et al. 2007). Therefore, there is a growing demand to differen- tiate, among the fermentative yeast flora, indigenous strains with typical oenological traits which could be considered representative of a particular oenological region. As the importance of the role of S. cerevisiae in winemaking has been established, the number of wine yeast strains available in the world market for use as starters has increased in the S. Orlic ´(&) Á T. Vojvoda Á K. H. Babic ´ Department of Microbiology, Faculty of Agriculture, University of Zagreb, Svetos ˇimunska 25, 10 000 Zagreb, Croatia e-mail: sorlic@agr.hr S. Orlic ´ Á F. N. Arroyo-Lo ´pez Institut ‘‘Cavanilles’’ De Biodiversitat i Biologia Evolutiva, Universitat de Vale `ncia, Edifici d’Instituts, Parc Cientı ´fic de Paterna, P.O. Box 22085, 46071 Valencia, Spain A. Jeromel Á B. Kozina Department of Viticulture and Enology, Faculty of Agriculture, University of Zagreb, Svetos ˇimunska 25, 10 000 Zagreb, Croatia L. Iacumin Á G. Comi Department of Food Science, Faculty of Agriculture, University of Udine, Via Sondrio 2, 33 100 Udine, Italy 123 World J Microbiol Biotechnol (2010) 26:1483–1489 DOI 10.1007/s11274-010-0323-9