Characterization of the recombinant Brettanomyces anomalus β-glucosidase and its potential for bioflavouring

Abstract

Aim: Plant materials used in the food industry contain up to five times more aromas bound to glucose (glucosides) than free, unbound aromas, making these bound aromas an unused flavouring potential. The aim of this study was to identify and purify a novel β-glucosidase from Brettanomyces yeasts that are capable of releasing bound aromas present in various food products. Methods and Results: We screened 428 different yeast strains for β-glucosidase activity and are the first to sequence the whole genome of two Brettanomyces yeasts (Brettanomyces anomalus and Brettanomyces bruxellensis) with exceptionally high β-glucosidase activity. Heterologous expression and purification of the identified B. anomalus β-glucosidase showed that it has an optimal activity at a higher pH (5·75) and lower temperature (37°C) than commercial β-glucosidases. Adding this B. anomalus β-glucosidase to cherry beers and forest fruit milks resulted in increased amounts of benzyl alcohol, eugenol, linalool and methyl salicylate compared to Aspergillus niger and Almond glucosidase. Conclusions: The newly identified B. anomalus β-glucosidase offers new possibilities for food bioflavouring. Significance and Impact of the Study: This study is the first to sequence the B. anomalus genome and to identify the β-glucosidase-encoding genes of two Brettanomyces species, and reports a new bioflavouring enzyme.