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Multiple integration of exo-β-1,3-glucanase (exgA) gene by using δ-sequence in Saccharomyces cerevisiae

초록

영어

β-1,3-glucanase was widely used in various biotechnological processes and overproduction was required for versatile industrial utilization. We subcloned β-1,3-glucanase gene (exgA) from Aspergillus oryzae and examined overexpression of β-1,3-glucanase by using δ -targeted multiple integration. We constructed two plasmids for multiple integration of exgA, pRSδ-exgA and pRSδH-exgA for first and second round integration, respectively. These plasmids contain ADH1 promoter and exoinulinase signal sequence (Inus.s) for secretion of β-1,3-glucanase. The pRSδ-exgA plasmid was transformed into S. cerevisiae BY4742△ exg1 strain. Subsequently, the pRSδH-exgA plasmid was transformed into S. cerevisiae BY4742△exg1/pRSδ-exgA strain. In the BY4742△exg1/pRSδ-exgA and BY4742△ exg1/pRSδ-exgA/pRSδH-exgA strains, β-1,3-glucanase activity was 0.94 and 1.4 unit/㎖�, respectively. Copy numbers of exgA gene were 6.7 and 10-fold increased by first and second round integration, respectively. These results suggested that exgA gene was successfully secreted and overexpressed by using δ-sequence integration. We will further increase of copy number by using repeated δ-integration.

저자정보

  • Min-Jung KIM Dept. of Biomaterial Control, Dong-Eui University, Busan 614-714, Korea.
  • Min-Ji KIM Dept. of Biotechnology and Bioengineering, Dong-Eui University, Busan 614-714, Korea.
  • Koichi TAMANO Reasearch Institute for Cell Engineering, National Institute of Advanced Industrial Science and Technology (AIST), Ibaraki 305-8566, Japan.
  • Masayuki MACHIDA Reasearch Institute for Cell Engineering, National Institute of Advanced Industrial Science and Technology (AIST), Ibaraki 305-8566, Japan.
  • Soo-Wan NAM Dept. of Biomaterial Control, Dong-Eui University, Busan 614-714, Korea. Dept. of Biotechnology and Bioengineering, Dong-Eui University, Busan 614-714, Korea.
  • Yeon-Hee KIM Dept. of Biomaterial Control, Dong-Eui University, Busan 614-714, Korea. Dept. of Biotechnology and Bioengineering, Dong-Eui University, Busan 614-714, Korea.

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