A β-glucosidase from Oenococcus oeni ATCC BAA-1163 with potential for aroma release in wine: cloning and expression in E. coli

Abstract

Lactic acid bacteria (LAB) are responsible for olfactory changes in wine during malolactic fermentation (MLF). A side characteristic of MLF is the release of grape derived aroma compounds from their glycosylated precursors by β-glycosidase activities of these bacteria. Apart from Oenococcus oeni, which is regarded as the most promising species for MLF, glycosidic activities have also been observed in wine related members of the genera Lactobacillus and Pediococcus. Nevertheless, information on the involved enzymes including their potential use in winemaking is limited. In this study we report that β-glucosidases with similar protein sequences can be identified in the genomes of Lactobacillus brevis, O. oeni and Leuconostoc mesenteroides. TTG serves as start codon for the glucosidase gene of O. oeni. The β-glucosidase of O. oeni ATCC BAA-1163 was expressed in E. coli and partially characterized. The enzyme displayed characteristics similar to β-glucosidases isolated from L. brevis and L. mesenteroides. A pH optimum between 5.0 and 5.5, and a K(m) of 0.17 mmol L(-1 )pNP-β-D-glucopyranoside were determined. A glycosyltransferase activity was observed in the presence of ethanol. The enzyme from O. oeni was capable to hydrolyze glycosides extracted from Muskat wine. This study also contains a report on glycosidase activities of several LAB species including Oenococcus kitaharae.

Identification and Localization of β-D-Glucosidase from Two Typical Oenococcus oeni Strains

“…However, the subcellular localization prediction showed the enzyme exists mainly in the cytoplasm as a soluble protein, which may be due to the secondary and tertiary structure of the protein that alters the hydrophobicity of βG. βG of the two strains was predicted as a non-secretory protein which mirrors the previous report that low βG activity was observed in the culture supernatant for both strains (Li et al, 2012b), as well as the subcellular localization prediction that little of the enzyme was extracellular.As for location of βG, intracellular enzyme has been reported for different O. oeni strains (Barbagallo et al, 2004;Michlmayr et al, 2010;Perez-Martin et al, 2012). The same results have also been observed for O. oeni SD-2a and 31MBR (Li et al, 2012b).…”

“…It has been well established that apart from free flavor compounds, a significant part of flavors remain in newly made wine as odorless non-volatile glycosides (Maicas and Mateo, 2005;Michlmayr et al, 2010). The odorless glycosides containing aroma and flavor aglycones are not directly accessible to the olfactory mucosa and may affect wine quality greatly after hydrolysis (Mesas et al, 2012;Michlmayr et al, 2012b).…”

“…βG activity in O. oeni, the main bacterial species conducting MLF, was confirmed more than 20 years ago (Guilloux-Benatier et al, 1993). Over the past decades, numerous investigations have been conducted, providing evidence for the potential βG activity of O. oeni strains for flavor enhancement in wines (Spano et al, 2005;Michlmayr et al, 2010;Gagné et al, 2011). It has been reported that the possession of glycosidic activities is widespread and strain dependent among O. oeni strains (Barbagallo et al, 2004;Grimaldi et al, 2005;Saguir et al, 2009).…”

Assessment of β-D-Glucosidase Activity from Two Typical Strains of the Lactic Acid Bacteria, Oenococcus oeni, in China

“…Recently, an increased interest in the sources of βG has been focused on lactic acid bacteria, especially Oenococcus oeni, the main bacterial species that conducts malolactic fermentation (MLF) in winemaking, as βG activity from grape and yeasts is limited in winemaking (Maicas & Mateo, 2005;Palmeri & Spagna, 2007;Saguir et al, 2009;Michlmayr et al, 2010b). Numerous investigations have been conducted of O. oeni strains, providing evidence of the potential βG activity for flavour enhancement in wines Boido et al, 2002;Mansfield et al, 2002;Ugliano et al, 2003;Barbagallo et al, 2004b;D'Incecco et al, 2004;Grimaldi et al, 2005a;Bloem et al, 2008;Michlmayr et al, 2010a;Gagné et al, 2011).…”

Assessment of β-d-glucosidase activity of intact cells of two Oenococcus oeni strains with synthetic and natural substrates

“…Thus the study of glycosidase from lactic acid bacteria, especially O. oeni strains, for aroma release during winemaking is of great interest. Over the past decades, numerous investigations have been conducted, providing evidence of the potential βG activity of O. oeni strains for flavour enhancement in (Michlmayr et al 2010a;Gagne et al 2011;Fatima et al 2012). Diverse O. oeni strains were tested with synthetic substrate to determine their capacity to release volatile compounds (McMahon et al 1999;Grimaldi et al 2000Grimaldi et al , 2005aGagne et al 2011;Michlmayr et al 2012a).…”