Characterization of a Thermophilic and Inhibitor-Tolerant GH1 β-Glucosidase Present in a Hot Spring

beta-glucosidase is a key enzyme in the degradation of lignocellulosic biomass, which is responsible for the conversion of oligosaccharides from cellulose hydrolysates to glucose. However, its required high temperatures and the presence of inhibitors have limited its use in industry. In this study, a new beta-glucosidase gene, named thbg2, was obtained from the metagenome Ruidian Hot Spring, Tengchong City, Yunnan Province, southwestern China. The gene was synthesized, cloned, heterologously expressed, and enzymatically characterized. Its optimum temperature and pH were 60 degrees C and pH 5.6, respectively. ThBg2 exhibited more than 60% relative activity in temperatures ranging from 40 degrees C to 70 degrees C and across a pH of 4.0-6.6. It maintained 100% relative activity after incubation at either 50 degrees C for 24 h or 60 degrees C for 12 h and more than 80% residual activity after incubation at pH 4.0-6.0 for 24 h. Moreover, it maintained more than 80% relative activity in the presence of heavy metal ions, ethanol, SDS etc. Furthermore, glucose yields from corn stalks increased by 20% after ThBg2 (0.05 mg/mL) was added to the commercial cellulase reaction system. Overall, this work identified a thermophilic and inhibitor-tolerant beta-glucosidase with potential applications in commercial lignocellulose utilization and the bioenergy industry.