Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Molecular Cloning and Expression of the Trichoderma harzianum C4 Endo-${\beta}-1$,4-Xylanase Gene in Saccharomyces cerevisiae

  • Lee, Jung-Min (Skin Biotechnology Center, Graduate School of Biotechnology and Institute of Life Science and Resources, Kyung Hee University) ;
  • Shin, Ji-Won (Skin Biotechnology Center, Graduate School of Biotechnology and Institute of Life Science and Resources, Kyung Hee University) ;
  • Nam, Jae-Kook (Skin Biotechnology Center, Graduate School of Biotechnology and Institute of Life Science and Resources, Kyung Hee University) ;
  • Choi, Ji-Young (School of Biological Sciences, University of Ulsan) ;
  • Jeong, Choon-Soo (School of Biological Sciences, University of Ulsan) ;
  • Han, In-Seob (School of Biological Sciences, University of Ulsan) ;
  • Nam, Soo-Wan (Department of Biotechnology and Bioengineering, Dong-Eui University) ;
  • Choi, Yun-Jaie (Department of Food and Animal Biotechnology, Seoul National University) ;
  • Chung, Dae-Kyun (Skin Biotechnology Center, Graduate School of Biotechnology and Institute of Life Science and Resources, Kyung Hee University)
  • Published : 2009.08.31
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Abstract

An endo-${\beta}-1$,4-xylanase (${\beta}$-xylanase) from Trichoderma harzianum C4 was purified without cellulase activity by sequential chromatographies. The specific activity of the purified enzyme preparation was 430 units/mg protein on D-xylan. The complementary DNA (cDNA) encoding ${\beta}$-xylanase (xynII) was amplified by PCR and isolated from cDNA PCR libraries constructed from T. harzianum C4. The nucleotide sequence of the cDNA fragment contained an open reading frame of 663 bp that encodes 221 amino acids, of which the mature protein is homologous to several ${\beta}$-xylanases II. An intron of 63 bp was identified in the genomic DNA sequence of xynII. This gene was expressed in Saccharomyces cerevisiae strains under the control of adh1 (alcohol dehydrogenase I) and pgk1 (phosphoglycerate kinase I) promoters in 2 ${\mu}$-based plasmids, which could render recombinants able to secrete ${\beta}$-xylanase into the media.

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