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Molecular Cloning and Expression of a Novel β-Glucosidase Gene from Phialophora sp. G5

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Abstract

A novel β-glucosidase gene, bgl1G5, was cloned from Phialophora sp. G5 and successfully expressed in Pichia pastoris. Sequence analysis indicated that the gene consists of a 1,431-bp open reading frame encoding a protein of 476 amino acids. The deduced amino acid sequence of bgl1G5 showed a high identity of 85 % with a characterized β-glucosidase from Humicola grisea of glycoside hydrolase family 1. Compared with other fungal counterparts, Bgl1G5 showed similar optimal activity at pH 6.0 and 50 °C and was stable at pH 5.0–9.0. Moreover, Bgl1G5 exhibited good thermostability at 50 °C (6 h half-life) and higher specific activity (54.9 U mg–1). The K m and V max values towards p-nitrophenyl β-d-glucopyranoside (pNPG) were 0.33 mM and 103.1 μmol min–1 mg–1, respectively. The substrate specificity assay showed that Bgl1G5 was highly active against pNPG, weak on p-nitrophenyl β-d-cellobioside (pNPC) and p-nitrophenyl-β-d-galactopyranoside (ONPG), and had no activity on cellobiose. This result indicated Bgl1G5 was a typical aryl β-glucosidase.

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Acknowledgments

This research was supported by the National Science and Technology Support Program (2011BADB02) and the National "948" Project (2011-G7-4) and the China Modern Agriculture Research System (CARS-42).

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Authors and Affiliations

  1. Key Laboratory of Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, No. 12 Zhongguancun South Street, Beijing, 100081, People’s Republic of China

    Xuejun Li, Junqi Zhao, Pengjun Shi, Peilong Yang, Yaru Wang, Huiying Luo & Bin Yao

  2. Beijing Challenge Bio-Technology Co., Ltd., Beijing, 100081, People’s Republic of China

    Xuejun Li

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  1. Xuejun Li
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  2. Junqi Zhao
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  3. Pengjun Shi
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  4. Peilong Yang
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  5. Yaru Wang
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  6. Huiying Luo
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  7. Bin Yao
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Correspondence to Bin Yao.

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Li, X., Zhao, J., Shi, P. et al. Molecular Cloning and Expression of a Novel β-Glucosidase Gene from Phialophora sp. G5. Appl Biochem Biotechnol 169, 941–949 (2013). https://doi.org/10.1007/s12010-012-0048-3

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  • DOI: https://doi.org/10.1007/s12010-012-0048-3

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