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Heterologous Expression and Characterization of a GH3 β-Glucosidase from Thermophilic Fungi Myceliophthora thermophila in Pichia pastoris

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Abstract

A novel β-glucosidase of glycoside hydrolase (GH) family 3 from Myceliophthora thermophila (mtbgl3b) was successfully expressed in Pichia pastoris. The full-length gene consists of 2613 bp nucleotides encoding a protein of 870 amino acids. MtBgl3b showed maximum activity at pH 5.0 and remained more than 70 % relative activity at 3.5–6.0. The enzyme displayed the highest activity at 60 °C and kept about 90 % relative activity for 50–65 °C; besides, the enzyme showed psychrophilic trait and remains 51 % relative activity at 40 °C. MtBgl3b exhibited good stability over a wide pH range of 3.0–10.0 and was thermostable at 60 and 65 °C. The enzyme displayed highest activity towards p-nitrophenyl-β-d-glucopyranoside (pNPG), followed by p-nitrophenyl-d-cellobioside (pNPC), cellotetraose, cellotriose, cellobiose, and gentiobiose. When using 10 % cellobiose (w/v) as the substrate, the enzyme showed transglycosylation activity to produce the cellotriose. The kinetic parametric of K m and V max were 2.78 mM and 927.9 μM mg−1 min−1, respectively. Finally, the reaction mode of the enzyme and the substrates were analyzed by molecular docking approach.

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Acknowledgments

This study was supported by the National High-Tech Research and Development Program (863 Program, No. 2012AA022203 and No. SS2014AA021301) and Tianjin Zhuanxiang Project (13ZCDZSY05000).

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  1. Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Xiqi Dao 32, Tianjin Airport Economic Area, Tianjin, 300308, China

    Junqi Zhao, Chao Guo, Chaoguang Tian & Yanhe Ma

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  1. Junqi Zhao
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Correspondence to Chaoguang Tian.

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Zhao, J., Guo, C., Tian, C. et al. Heterologous Expression and Characterization of a GH3 β-Glucosidase from Thermophilic Fungi Myceliophthora thermophila in Pichia pastoris . Appl Biochem Biotechnol 177, 511–527 (2015). https://doi.org/10.1007/s12010-015-1759-z

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