Cloning and expression of an Aspergillus kawachii endo-1,4-β-xylanase gene in Saccharomyces cerevisiae

Johan M. Crous, Isak S. Pretorius, Willem H. van Zyl*

*Corresponding author for this work

Research output: Contribution to journalArticle

55 Citations (Scopus)

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 (PGK1p) and terminator (PGK1T) sequences. The PGK1P-xynC-PGK1T 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 pH3, 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.

Original languageEnglish
Pages (from-to)467-473
Number of pages7
JournalCurrent Genetics
Volume28
Issue number5
DOIs
Publication statusPublished - Oct 1995
Externally publishedYes

Fingerprint Dive into the research topics of 'Cloning and expression of an Aspergillus kawachii endo-1,4-β-xylanase gene in Saccharomyces cerevisiae'. Together they form a unique fingerprint.

  • Cite this