High-level expression of a novel Penicillium endo-1,3(4)-β-D-glucanase with high specific activity in Pichia pastoris.

Journal of Industrial Microbiology & Biotechnology
Xiaoyan ChenBin Yao

Abstract

A novel endo-1,3(4)-β-D-glucanase gene (bgl16C1) from Penicillium pinophilum C1 was cloned and sequenced. The 945-bp full-length gene encoded a 315-residue polypeptide consisting of a putative signal peptide of 18 residues and a catalytic domain belonging to glycosyl hydrolase family 16. The deduced amino acid sequence showed the highest identity (82%) with the putative endo-1,3(4)-β-glucanase from Talaromyces stipitatus ATCC 10500 and 60% identity with the characterized β-1,3(4)-glucanase from Paecilomyces sp. FLH30. The gene was successfully overexpressed in Pichia pastoris. Recombinant Bgl16C1 constituted 95% of total secreted proteins (2.61 g l⁻¹) with activity of 28,721 U ml⁻¹ in a 15-l fermentor. The purified recombinant Bgl16C1 had higher specific activity toward barley β-glucan (12,622 U mg⁻¹) than all known glucanases and also showed activity against lichenan and laminarin. The enzyme was optimally active at pH 5.0 and 55°C and exhibited good stability over a broad acid and alkaline pH range (>85% activity at pH 3.0-7.0 and even 30% at pH 11.0). All these favorable enzymatic properties make it attractive for potential applications in various industries.

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Citations

Apr 16, 2016·AMB Express·Ghulam MustafaAmer Jamil
Feb 18, 2015·Journal of Biotechnology·Sebastian C SpohnerPeter Czermak
Feb 15, 2020·Journal of Cellular Physiology·Mohsen KarbalaeiHadi Farsiani
Jan 13, 2021·Bioresource Technology·Juan A Méndez-LíterMaría Jesús Martínez
Oct 1, 2018·International Journal of Biological Macromolecules·Barkat AliHai Zhao
Jul 26, 2018·Journal of Agricultural and Food Chemistry·Shuai YouHui-Ying Luo

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