SCREENING OF ENZYMATIC ACTIVITIES IN NON-SACCHAROMYCESCIDER YEASTS

Abstract

The activities of polygalacturonase, pectin lyase, β‐glucosidase, β‐xylosidase and protease were determined using solid media in 420 wild non‐Saccharomyces cider strains identified by internal transcribed spacer‐restriction fragment length polymorphism. The identified species corresponded to Hanseniaspora valbyensis, Hanseniaspora uvarum, Hanseniaspora osmophila, Metschnikowia pulcherrima, Candida parapsilosis and Pichia guilliermondii. The most common activity exhibited was that of β‐glucosidase (33%), with all the analyzed species having some strains able to develop this activity. Strains of M. pulcherrima showed the greatest capacity to produce β‐glucosidase and protease. β‐xylosidase was detected in 17 yeast strains belonging to the genera Hanseniaspora, Pichia and Metschnikowia. All of the tested species have some strains with the capacity to develop β‐xylosidase activity, except for C. parapsilosis. No strains were able to secrete pectin lyase, while polygalacturonase activity was observed in eight Hanseniaspora strains. Only two strains, belonging to the species H. uvarum and M. pulcherrima, developed three enzymatic activities, namely β‐glucosidase, β‐xylosidase and protease. PRACTICAL APPLICATIONS The non‐Saccharomyces yeasts are very interesting in cider making because of the low level of ethanol in cider. In this paper, it is demonstrated that the yeast isolated from Asturian cider represents a source of several enzymes, which have capacity to release flavor compounds. In this sense non‐Saccharomyces strains are being used as inoculated cultures to ferment apple pomace and apple must, with the aim of enhancing the aroma and flavor of the products.

Hanseniaspora uvarum from Winemaking Environments Show Spatial and Temporal Genetic Clustering

“…Hanseniaspora uvarum (anamorph Kloeckera apiculata ) is an apiculate yeast species frequently found on mature fruits (Spencer et al, 1992 ; Morais et al, 1995 ) and particularly on grapes where it forms part of the grape and fermentation microbiome.Its association with grapes and the first stages of alcoholic fermentation (AF) has been reported repeatedly during the last century (Castelli, 1955 ; Schütz and Gafner, 1993 ; Hierro et al, 2006 ) and for most—if not all—vineyard regions worldwide (Heard and Fleet, 1985 ; Holloway et al, 1990 ; Mateo et al, 1991 ; Comi et al, 2001 ; Beltran et al, 2002 ; Jolly et al, 2003 ; Combina et al, 2005 ; Li et al, 2010 ; Zott et al, 2010 ; Kachalkin et al, 2015 ). H. uvarum is also frequently isolated from other fermented beverages such as cider (Lachance, 1995 ; Cabranes et al, 1997 ; Valles et al, 2007 ; Pando Bedrinana et al, 2012 ), palm wine and cashew juice (Owuama and Saunders, 1990 ), tequila (Bilbao et al, 1997 ), sugar-cane aguardente (Morais et al, 1997 ), etc. It is part of the natural microbiome of many fermented food processes, including coffee (Masoud et al, 2004 ) and cocoa (Batista et al, 2015 ) fermentations.…”

Characterization of tolerance and multi‐enzyme activities in non‐Saccharomycesyeasts isolated from Vidal blanc icewine fermentation

“…For example, the β‐glucosidase was not detected in T. delbrueckii HB02, HB03, HC04, and HG01, S. bacillaris HA01 and HB07, and D. hansenii HC01, which may be due to the differences between species or the lack of a complex growth medium and the fastidious nature of the organisms. Furthermore, it have been previously reported that the ability of wine yeasts to produce β‐xylosidase was very rare and was currently only found in the genera Hanseniaspora , Pichia Metschnikowia , Torulaspora, and Candida (Bedriñana et al, ; Bokulich et al, ; Manzanares, Ramón, & Querol, ; Mateo et al, ). Interestingly, the β‐xylosidase activity was detected in D. hansenii HB08 in this study.…”

“…The release of volatile aroma is related to hydrolytic enzymes secreted by non-Saccharomyces yeasts, with β-glucosidase, β-xylosidase, and pectinase having outstanding contributions (Belda et al, 2016;Canonico, Agarbati, Comitini, & Ciani, 2015;Comitini et al, 2011;López et al, 2015). In previous studies, strains in the genera Candida, Debaryomyces, Hanseniaspora, Metschnikowia, and Torulaspora were found to secrete β-glucosidase (Bedriñana et al, 2012;Fernández, Ubeda, & Briones, 2000;Iranzo, Otero, & Brionesperez, 2010;Mateo et al, 2011). However, in the same species, strains produced different enzymatic activities.…”

“…Non‐Saccharomyces yeasts can secrete several beneficial hydrolytic enzymes (such as β‐glucosidase, β‐xylosidase, and pectinase) which are absent in Saccharomyces yeasts (Andorrà, María, Mas, Esteve‐Zarzoso, & Guillamón, ; Zhang, Shen, Duan, & Yan, ). These enzymes provide interesting wine organoleptic characteristics (Bedriñana, Queipo, & Valles, ). β‐Glucosidase and β‐xylosidase are involved in the release of terpenes, which is beneficial for improving aromatic complexity of wine (López, Mateo, & Maicas, ).…”

Advances in the Study of Candida stellata

“…The Saccharomyces genus is not considered as a good producer of exogenous enzymes. Instead, several non-Saccharomyces yeast species exhibit natural enzymatic activities [91]. The enzymes of interest produced by these yeasts include esterases, lipases, glycosidases, proteases, and cellulases usually related to hydrolysis of structural components [4,92].…”