β-1, 4-glucosidases generate glucose from cellobiose and oligosaccharides, enhancing the productivity in biorefinery and the bioconversion process as well as the nutritional value in food and feed. With the high-throughput sequencing technique, a novel β-1, 4-glucosidase, named bgl T2, containing 861 amino acid residues, was found from Aspergillus fresenii. bgl T2 belongs to the glycosyl hydrolase (GH) family 3. The bgl T2 that expressed by Komagataella phaffii X33 presented the highest activity at 55 °C and pH 5.5. The half-lives of bgl T2 under 50 °C, 55 °C, 60 °C, and 65 °C were 9 min 36 s, 4 min 22 s, 117 s, and 68 s, respectively. The bgl T2 was stable between pH 3.0 to pH 8.0. The Michaelis constant (K m) and the theoretical maximum rate (V max) of bgl T2 were 0.0007 mol/L and 9 × 10-8 mol/L/s, respectively. In a 5 L fermentation vessel, the recombinant K.phaffii X33 could yield a β-1, 4-glucosidase activity of 4.45 U/mL after 96 h methanol inducement. As an important member of cellulases, the novel bgl T2 might contribute to bioenergy, food processing, feed enrichment, and nutritional study, etc. This study also developed a path to obtain new enzymes depending on high-throughput sequencing technique.

中文翻译：

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异源表达的新型β-1，4-葡糖苷酶的酶学特征来源于费氏曲霉。

β-1，4-葡萄糖苷酶从纤维二糖和寡糖中产生葡萄糖，从而提高了生物精炼厂和生物转化过程的生产率以及食品和饲料的营养价值。利用高通量测序技术，从费氏曲霉中发现了一种新的β-1、4-葡糖苷酶，名为bgl T2，含有861个氨基酸残基。bgl T2属于糖基水解酶（GH）家族3。由Komagataella phaffii X33表达的bgl T2在55°C和pH 5.5时表现出最高的活性。bgl T2在50°C，55°C，60°C和65°C下的半衰期分别为9分钟36 s，4分钟22 s，117 s和68 s。bgl T2在pH 3.0至pH 8.0之间稳定。bgl T2的米氏常数（K m）和理论最大速率（V max）分别为0.0007 mol / L和9×10-8 mol / L / s。在5升发酵罐中 甲醇诱导96 h后，重组K.phaffii X33可产生4.45 U / mL的β-1，4-葡萄糖苷酶活性。作为纤维素酶的重要成员，新型bgl T2可能有助于生物能源，食品加工，饲料富集和营养研究等。这项研究还开发了一种依靠高通量测序技术获得新酶的途径。