The enzymatic characters of heterologous expressed novel β-1, 4-glucosidase originated from .

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 T2, containing 861 amino acid residues, was found from . T2 belongs to the glycosyl hydrolase (GH) family 3. The T2 that expressed by X33 presented the highest activity at 55 °C and pH 5.5. The half-lives of 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 T2 was stable between pH 3.0 to pH 8.0. The Michaelis constant ( ) and the theoretical maximum rate ( ) of T2 were 0.0007 mol/L and 9 × 10 mol/L/s, respectively. In a 5 L fermentation vessel, the recombinant 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 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.