Cloning and expression of an Aspergillus kawachii endo-1,4-β-xylanase gene in Saccharomyces cerevisiae
Date
1995
Authors
Crous J.M.
Pretorius I.S.
Van Zyl W.H.
Journal Title
Journal ISSN
Volume Title
Publisher
Abstract
First-strand cDNA was prepared from mRNA isolated from Aspergillus kawachii IFO4308 and the β-xylanase gene (xynC) amplified by using the polymerase chain reaction (PCR) technique. This gene was inserted between the yeast phosphoglycerate kinase (PGK1) gene promoter (PGK1(p)) and terminator (PGK1(T)) sequences. The PGK1(p)-xynC-PGK1(T) construct (designated XYN3) was cloned into a multicopy episomal plasmid and the XYN3 gene was expressed in Saccharomyces cerevisiae. Functional β-xylanase (Xyn3) was produced and secreted by the recombinant yeast. Xyn3 was stable between 30 and 50°C, and the optimum temperature and pH were shown to be at 60°C and lower than pH 3, respectively. An autoselective fur1::LEU2 XYN3 recombinant strain was developed that allowed β-xylanase production at a level of 300 nkat/ml in a non-selective complex medium.
Description
Keywords
complementary dna, fungal rna, messenger rna, phosphoglycerate kinase, recombinant dna, xylan endo 1,3 beta xylosidase, article, aspergillus, enzyme release, enzyme stability, enzyme synthesis, gene expression, gene insertion, molecular cloning, nonhuman, ph, plasmid, polymerase chain reaction, priority journal, promoter region, saccharomyces cerevisiae, thermostability, Aspergillus, Base Sequence, Cloning, Molecular, DNA, Fungal, Molecular Sequence Data, Saccharomyces cerevisiae, Support, Non-U.S. Gov't, Xylosidases, Aspergillus, Aspergillus kawachii, Saccharomyces cerevisiae
Citation
Current Genetics
28
5
28
5