High-level expression of a specific beta-1,3-1,4-glucanase from the thermophilic fungus Paecilomyces thermophila in Pichia pastoris

Applied Microbiology and Biotechnology
Chengwei HuaPriti Katrolia

Abstract

In this study, a novel beta-1,3-1,4-glucanase gene (designated as PtLic16A) from Paecilomyces thermophila was cloned and sequenced. PtLic16A has an open reading frame of 945 bp, encoding 314 amino acids. The deduced amino acid sequence shares the highest identity (61%) with the putative endo-1,3(4)-beta-glucanase from Neosartorya fischeri NRRL 181. PtLic16A was cloned into a vector pPIC9K and was expressed successfully in Pichia pastoris as active extracellular beta-1,3-1,4-glucanase. The recombinant beta-1,3-1,4-glucanase (PtLic16A) was secreted predominantly into the medium which comprised up to 85% of the total extracellular proteins and reached a protein concentration of 9.1 g l(-1) with an activity of 55,300 U ml(-1) in 5-l fermentor culture. The enzyme was then purified using two steps, ion exchange chromatography, and gel filtration chromatography. The purified enzyme had a molecular mass of 38.5 kDa on SDS-PAGE. It was optimally active at pH 7.0 and a temperature of 70 degrees C. Furthermore, the enzyme exhibited strict specificity for beta-1,3-1,4-D: -glucans. This is the first report on the cloning and expression of a beta-1,3-1,4-glucanase gene from Paecilomyces sp.

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Citations

Dec 16, 2011·Applied Microbiology and Biotechnology·Ya-Shan ChengRey-Ting Guo
Feb 23, 2012·Journal of Industrial Microbiology & Biotechnology·Xiaoyan ChenBin Yao
Apr 12, 2012·Applied and Environmental Microbiology·Yu-Lung HungYo-Chia Chen
Nov 23, 2013·Biochimica Et Biophysica Acta·Ya-Shan ChengRey-Ting Guo
Jan 10, 2016·World Journal of Microbiology & Biotechnology·Ting XuShujiang Li
Oct 27, 2015·Bioprocess and Biosystems Engineering·Burcu Gündüz Ergün, Pınar Çalık
Oct 3, 2014·Bioscience, Biotechnology, and Biochemistry·Lerato Mary SekhoholaAshton Keith Cowan
Jan 9, 2014·Biotechnology and Applied Biochemistry·Zhen Gao
Nov 16, 2018·Journal of the Science of Food and Agriculture·Fatma Chaari, Semia Ellouz Chaabouni
Dec 14, 2016·The Journal of Biological Chemistry·Zhen QinZhengqiang Jiang
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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