Heterologous expression of the Bacillus pumilus endo-?-xylanase (xynA) gene in the yeast Saccharomyces cerevisiae

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• 作者：Filip Nuyens ; Willem H. van Zyl ; Dirk Iserentant ; Hubert Verachtert and Chris Michiels
• 刊名：Applied Microbiology and Biotechnology
• 出版年：2001
• 出版时间：August 2001
• 年：2001
• 卷：56
• 期：3-4
• 页码：431-434
• 全文大小：61 KB
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Biotechnology
  Microbiology
  Microbial Genetics and Genomics
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1432-0614

文摘

The endo-#-xylanase-encoding gene (xynA) of Bacillus pumilus PLS was isolated from a genomic DNA library and the open reading frame (ORF) was inserted in expression vectors for the yeast Saccharomyces cerevisiae. Plasmid pFN3 harboured the xynA ORF fused to the yeast mating pheromone alpha-factor signal sequence (MFŋS) under the control of the alcohol dehydrogenase II gene promotor (ADH2P) and terminator (ADH2T) sequences. In plasmid pFN4, the MFŋS-xynA ORF was brought under the control of the phosphoglycerate kinase I gene promotor (PGK1P) and terminator (PGK1T) sequences. Autoselective, recombinant S. cerevisiae [fur1::LEU2] strains bearing pFN3 or pFN4 secreted functional endo-#-xylanase when grown in complex medium. Enzymatic activities in the culture supernatants reached maximum levels of 8.5 nkat/ml and 4.5 nkat/ml, respectively. The temperature and pH optimum for both the bacterial and the recombinant xylanase were 58 °C and pH 6.2.