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β-glucosidases have been widely used in food, medicine, bioenergy and other fields, and thus it is necessary to explore new and efficient β-glucosidases.[Objective] To realize the prokaryotic expression of a GH1 glucosidase derived from Devosia psychrophila and characterize the enzymatic properties of the expressed protein. [Methods] The gene encoding the β-glucosidase derived from D. psychrophila was synthesized, named bgl59, and then transformed into Escherichia coli BL21(DE3). After the gene expression was induced, and the obtained protein was purified and characterized for the enzymatic properties. [Results] Bgl59 had a molecular weight of 48.8 kDa, with the highest activity at 55 ℃ and pH 6.0. After treatment for 1 h within the range of pH 5.0–8.5, Bgl59 maintained the relative activity over 80%. Bgl59 had the highest hydrolysis ability for 4-nitrophenyl-β-D-glucopyranoside (pNPG) among the eight substrates tested, with the Km of 3.090 mmol/L, Vmax of 194 μmol/(min·mg), and kcat of 159 s−1. The presence of 1 mmol/L of Ca2+ and Co2+ had a significant activating effect on Bgl59, while the presence of 0.1% SDS resulted in a complete loss of enzyme activity. The presence of 0.10 mol/L glucose and 0.30 mol/L xylose increased the activity of Bgl59 by 74% and 91%, respectively. Moreover, the enzyme remained the relative activity above 50% even when being cultured with 1.25 mol/L glucose or 2.00 mol/L xylose. [Conclusion] Bgl59 exhibits outstanding enzymatic properties, robust pH stability, and tolerance to metal ions and chemical reagents. It is a rare glucose-activated β-glucosidase with exceptional tolerance to glucose, holding significant potential for future industrial production and application.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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