Secretive expression of bacterial β-xylosidase gene including hexahistidin-tag in Pichia pastoris

How to Cite:Shirin YousefianOmid Ranaei siadatEhsan DehnaviMohammadtaghy BarChianBorojeniFarnaz Nikzad jamnaniSecretive expression of bacterial β-xylosidase gene including hexahistidin-tag in Pichia pastoris.koomesh.14(4):e152592.

Abstract

Introduction: β-D-Xylosidases (EC 3.2.1.37), one of the most important hemicellulases with high potential industrial application such as bioethanol production, are exo-type glycosidases that catalyze the successive removal of β-xylosyl residues from the non-reducing termini of xylobiose and higher linear β-1,4-xylooligosaccharides and in conjunction with β-xylanases are essential in completely depolymerizing xylan. The aim of this study was secretive expression of bacterial β-xylosidase gene including hexahistidin-tag in Pichia pastoris in order to achieve high level expression and enzyme purification subsequently. Materials and Methods: In this study, after optimizing the sequence of protein encoding bacterial β-xylosidase based on the expression codon usage of Pichia pastoris and addition of 6xHis-tag sequence through suitable designed primers to gene, transformation of recombinant plasmids was performed through electroporation method into the expression yeast. Results: The results showed that the recombinant β-xylosidase production was 40 U/L with 0/6 U/mg specific activity in flask culture. Conclusion: Cloning and successful expression of bacterial β-xylosidase gene was carried out in order to obtain an active and stable enzyme with high level expression from yeast expression system

Keywords

Yeasts, Xylosidases, Histidine, Gene expressio

مخمرها، گزایلوزیدازها، هیستیدین، بیان ژنی

comments