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Heterologous expression and purification of protopectinase-N from Bacillus subtilis in Pichia pastoris

Paper ID Volume ID Publish Year Pages File Format Full-Text
36635 45138 2006 5 PDF Available
Title
Heterologous expression and purification of protopectinase-N from Bacillus subtilis in Pichia pastoris
Abstract

A eukaryotic expression system for secretory production of the protopectinase from Bacillus subtilis XZ2 in Pichia Pastoris was constructed in this paper. The protopectinase gene without native signal sequence was amplified by PCR and fused to the pPICZαA plasmid containing α-factor sequence encoding secretion signal from Saccharomyces cerevisiae. The heterologous gene was expressed under the AOX1 promoter in a culture of P. pastoris X-33 in flasks. The recombinant protein was further purified by ammonium sulfate precipitate, Sephadex G75 and ion exchange chromatography and resulted in 18.91-fold purification; the purified protein was a molecular weight about 43 kDa and had an activity of 1948.64 U/mg.

Keywords
Bacillus subtilis; Heterologous expression; Protopectinase-N; Purification; Pichia pastoris
First Page Preview
Heterologous expression and purification of protopectinase-N from Bacillus subtilis in Pichia pastoris
Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 41, Issue 4, April 2006, Pages 975–979
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering