To obtain yeast strains with high β-glucosidase productivity and excellent tolerance to wine habitat, this study screened 452 native non-Saccharomyces cerevisiae in esculin culture medium, after which their fermentability, tolerance, β-glucosidase productivity, and enzymatic properties were analyzed. The results revealed 221 strains with CO2 loss greater than 0.51 g/100mL for 72 h, as well as 34 strains with high β-glucosidase productivity. Furthermore, GC204, NM218, ZC278, ZC287, BF345, and BF370 showed higher tolerance to glucose, SO2 (250 mg/L), ethanol (3%-6%), pH (2.5), and high sugar levels (350 g/L), respectively. Meanwhile, the above six yeast strains were further screened for high-yield of β-glucosidase, where GC204, NM218, and BF345 strains showed high enzyme activity of 54.34, 49.5, and 46.42 mU/mL, respectively. Further investigation of the enzymatic properties revealed NM218 and BF345 to have optimum reaction temperature and pH value for β-glucosidase production at 40 ℃ and 4.0, respectively. Also, Zn2+, Mn2+, Fe2+, and Fe3+ at a concentration of 5 mmol/L and cellobiose had a significant activation effect on the β-glucosidase activity of both strains whereby cellobiose increased the enzyme activity by 30.18%-40.80%. Contrary, Cu2+ had an inhibitory effect on β-glucosidase activity. Additionally, a 99% homology was obtained by 26S rDNA D1/D2 sequencing comparison where NM218 and BF345 were identified as Meyerozyma guilliermondii and Hanseniaspora uvarum, respectively.
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