Ochratoxin A (OTA) is a well-known, natural contaminant in foods and feeds because of its toxic effects, such as nephrotoxicity in various animals. Recent studies have revealed that Alcaligenes faecalis could generate enzymes to efficiently degrade OTA to ochratoxin α (OTα) in vitro. In an effort to obtain the OTA degrading mechanism, we purified and identified a novel degrading enzyme, N-acyl-L-amino acid amidohydrolase (AfOTase), from A. faecalis DSM 16503 via mass spectrometry. The same gene of the enzyme was also encountered in other A. faecalis strains. AfOTase belongs to peptidase family M20 and contains metal ions at the active site. In this study, recombination AfOTase was expressed and characterized in Escherichia coli. The molecular mass of recombinant rAfOTase was approximately 47.0 kDa, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme exhibited a wide temperature range (30–70 °C) and pH adaptation (4.5–9.0) and the optimal temperature and pH were 50 °C and 6.5, respectively.

The application of β-glucosidase from non-Saccharomyces yeasts to improve wine aroma has been widely explored. However, few enzymes are active under the severe conditions of wine aging (high ethanol concentration, low temperature, and low pH). Therefore, the application of β-glucosidase in wine aging needs further research. In this study, the β-glucosidases Mg-βgl and Hu-βgl extracted from Meyerozyma guilliermondii NM218 and Hanseniaspora uvarum BF345 were purified and used in young Chardonnay wines aged for 50 days. The enzyme activity of the two enzymes was measured. The effects of the two enzymes and a commercial β-glucosidase (An-βgl) on the volatile composition and sensory quality of the wine were also determined. The results showed that Mg-βgl and Hu-βgl had high specific activity of 1.95 U/mg and 2.11 U/mg, respectively, maintaining the activity of 70–80% at 20 °C, pH of 3.0–4.0, and 15% ethanol, corresponding to wine aging conditions. Analysis of volatiles with GC-MS showed a 65–70% increase in total terpenoids and new detection of C₁₃-norisoprenoids when the wines were treated with the three β-glucosidases. In addition, wines treated with Mg-βgl and Hu-βgl had more hexanol, phenylethanol, ethyl octanoate, ethyl heptanoate, and ethyl caprate than wines treated without and with An-βgl. In sensory analysis, the judges showed a greater preference for Hu-βgl-treated wines, to which they attributed pleasant sweet, floral, honey, pomelo, and banana aromas. The results of this study not only offer a way to improve flavor complexity in wine but also provide a reference for the use of other edible sources of β-glucosidase in wine aging.