ORIGINAL PAPER Characterization of b-glucosidases from Hanseniaspora sp. and Pichia anomala with potentially aroma-enhancing capabilities in juice and wine Jutaporn Swangkeaw • Sukanda Vichitphan • Christian E. Butzke • Kanit Vichitphan Received: 29 September 2009 / Accepted: 5 June 2010 / Published online: 12 June 2010 Ó Springer Science+Business Media B.V. 2010 Abstract The properties of intracellular b-glucosidases produced from two yeast isolates identified as Hansenias- pora sp. BC9 and Pichia anomala MDD24 were charac- terized. b-Glucosidase from Hanseniaspora sp. BC9 was not inhibited by both 20% w/v fructose and 20% w/v sucrose and was slightly inhibited by glucose ( [ 40% relative b-glucosidase activity with 10% w/v glucose). b-Glucosidase from P. anomala MDD24 was inhibited by glucose, fructose and sucrose. In the presence of 4–12% v/v ethanol, b-glucosidase from P. anomala MDD24 was stimulated in range 110–130% relative activity whereas b-glucosidase from Hanseniaspora sp. BC9 was substan- tially inhibited in the presence of ethanol. Finally, juice and wine of the Muscat-type grape variety, Traminette, were selected to determine sugar-bound volatile aroma release, particularly terpenes, by the activity of those b-glucosi- dases. The results showed that high concentration of free aroma compounds were detected from Traminette juice treated with b-glucosidase from Hanseniaspora sp. BC9 and Traminette wine treated with b-glucosidase from P. anomala MDD24. The preliminary results with pro- posed an application of these enzymes in commercial wine production lead to more efficient of b-glucosidase from Hanseniaspora sp. BC9 in releasing desirable aromas during an early stage of alcoholic fermentation while b-glucosidase from P. anomala MDD24 is suitable at the final stage of alcoholic fermentation. Keywords b-Glucosidase  Hanseniaspora sp.  Pichia anomala  Monoterpenes  Wine aroma Introduction Aroma by nose is the most important characteristic of wine contributing to its perceived quality and value (Armanino et al. 2008). Wine aroma is the outcome of a complex interaction between components present in the grape, those produced during fermentation, and those arising during aging (Hernandez et al. 2002). Many precursors of aro- matic components of juices, musts, wines such as the monoterpenes (limonene, linalool oxide, linalool, geraniol, nerol, citronellol, a-terpineol, etc.) are initially present in di-glycosidically-bound and non-volatile forms. These contain b-D-glucopyranoside bound directly to an aglycone and/or other sugars such as a-L-rhamnopyranose and a-L-arabinofuranose (Wilson et al. 1986; Vorin et al. 1990; Wu et al. 1991; Spagna et al. 2002a). Traditionally, these aromas are gradually released by acid hydrolysis during the aging of wine at typical cellar temperature around 12–15°C. Accelerated aging via low pH and high temper- ature conditions can be used for the hydrolysis of the bound aroma fraction, but may lead to extensive rearrangements of volatiles and the loss of the natural varietals aroma J. Swangkeaw Department of Microbiology, Faculty of Science, Khon Kaen University, Khon Kaen 4002, Thailand S. Vichitphan  K. Vichitphan (&) Department of Biotechnology, Faculty of Technology, Khon Kaen University, Khon Kaen 40002, Thailand e-mail: kanvic@kku.ac.th S. Vichitphan  K. Vichitphan Fermentation Research Center for Value Added Agricultural Products (FerVAAP), Khon Kaen University, Khon Kaen 40002, Thailand C. E. Butzke Department of Food Science, Purdue University, West Lafayette, IN 47907, USA 123 World J Microbiol Biotechnol (2011) 27:423–430 DOI 10.1007/s11274-010-0474-8