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http://dx.doi.org/10.4014/mbl.1906.06008

Optimal Expression System for Production of Recombinant Neoagarobiose Hydrolyase in Saccharomyces cerevisiae  

Jung, Hye-Won (Biomedical Engineering and Biotechnology Major, Division of Applied Bioengineering, Dong-Eui University)
Kim, Yeon-Hee (Biomedical Engineering and Biotechnology Major, Division of Applied Bioengineering, Dong-Eui University)
Publication Information
Microbiology and Biotechnology Letters / v.47, no.4, 2019 , pp. 662-666 More about this Journal
Abstract
In this study, the NABH558 gene expression system was constructed to efficiently produce neoagarobiose hydrolase (NABH) in Saccharomyces cerevisiae strain. The ADH1 and GAL10 promoters of the pAMFα-NABH and pGMFα-NABH plasmids were examined to determine the suitable promoter for the NABH558 gene expression, respectively. The effect of promoter and carbon sources on NABH558 gene expression was investigated by transforming each plasmid into the S. cerevisiae 2805 strain. The NABH activity in the 2805/pAMFα-NABH strain was 0.069 unit/ml/DCW in YPD medium, whereas that in the 2805/pGMFα-NABH strain was similar (0.02-0.027 unit/ml/DCW) irrespective of the medium composition. The higher NABH activity in the YPD medium was due to the increased NABH558 gene transcription. NABH produced in the recombinant strains could degrade agarose to galactose and AHG. This indicated that ADH1 promoter was a more optimal promoter for the expression of NABH558 gene than the GAL10 promoter. The NABH activity induced by the ADH1 promoter was about 3-fold higher than that induced by the GAL10 promoter.
Keywords
Carbon source; neoagarobiose hydrolase; promoter strength; Saccharomyces cerevisiae; signal sequence;
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