Purification and biochemical characterization of a novel thermophilic exo-β-1,3-glucanase from the thermophile biomass-degrading fungus Thielavia terrestris Co3Bag1

Background: The aim of this work was to purify and characterize exo-β-1,3-glucanase, namely, TtBgnA, from the thermophilic fungus Thielavia terrestris Co3Bag1 and to identify the purified enzyme. Results: The thermophilic biomass-degrading fungus T. terrestris Co3Bag1 displayed β-1,3-glucanase activity when grown on 1% glucose. An exo-β-1,3-glucanase, with an estimated molecular mass of 129 kDa, named TtBgnA, was purified from culture filtrates from T. terrestris Co3Bag1. The enzyme exhibited optimum activity at pH 6.0 and 70°C and half-lives (t1/2) of 54 and 37 min at 50 and 60°C, respectively. Substrate specificity analysis showed that laminarin was the best substrate studied for TtBgnA. When laminarin was used as the substrate, the apparent KM and Vmax values were determined to be 2.2 mg mL-1 and 10.8 U/mg, respectively. Analysis of hydrolysis products by thin-layer chromatography (TLC) revealed that TtBgnA displays an exo mode of action. Additionally, the enzyme was partially sequenced by tandem mass spectrometry (MS/MS), and the results suggested that TtBgnA from T. terrestris Co3Bag1 could be classified as a member of the GH-31 family. Conclusions: This report thus describes the purification and characterization of TtBgnA, a novel exo-β-1,3-glucanase of the GH-31 family from the thermophilic fungus T. terrestris Co3Bag1. Based on the biochemical properties displayed by TtBgnA, the enzyme could be considered as a candidate for potential biotechnological applications.How to cite: Rodríguez-Mendoza J, Santiago-Hernández A, Alvarez-Zúñiga MT, et al. Purification and biochemical characterization of a novel thermophilic exo-β-1,3-glucanase from the thermophile biomass-degrading fungus Thielavia terrestris Co3Bag1. Electron J Biotechnol 2019;41. https://doi.org/10.1016/j.ejbt.2019.07.001 Keywords: Characterization exo-glucanase, Exo-β-1,3-glucanase, Fungal glucanases, GH31 family, Laminarin, Purification exo-glucanase, Tandem mass spectrometry, Thermophilic enzyme, Thermophilic fungus, Thielavia terrestris Co3Bag1, TtBgnA