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KMID : 0613820130230070863
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Journal of Life Science
2013 Volume.23 No. 7 p.863 ~ p.868
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Efficient Secretory Expression of Recombinant Endoxylanase from Bacillus sp. HY-20 in Saccharomyces cerevisiae
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Kim Min-Ji
Kim Bo-Hyun
Nam Soo-Wan
Choi Eui-Sung
Shin Dong-Ha
Cho Han-Young
Son Kwang-Hee
Park Ho-Yong
Kim Yeon-Hee
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Abstract
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The XylP gene, which encodes endoxylanase in Bacillus sp. HY-20, was subcloned, and two expression plasmids, pG-xylP and pGMF-xylP were constructed. These plasmids, which contain different signal sequences, XylP s.s and MFα_opt s.s, respectively, for the secretory expression of endoxylanase, were transformed into Saccharomyces cerevisiae SEY2102 and FY833, respectively. The recombinant endoxylanases were successfully expressed, with a total activity range of 23.7-70.1 unit/ml according to the expression system and host strain. The endoxylanase activity in SEY2102/pGMF-xylP reached a maximum of 88.1 unit/ml in baffled flask culture. Most of the recombinant endoxylanase was efficiently secreted in the extracellular fraction, and the MFα_opt s.s was more efficient for secreting endoxylanase in yeast than the XylP s.s. Therefore, the expression system developed in this study produces large extracellular amounts of endoxylanase using S. cerevisiae as the host strain, and it could be used in bioethanol production and industrial applications.
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KEYWORD
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Endoxylanase, optimized MFα signal sequence (MFα_opt s.s), GAL promoter, secretory production, Saccharomyces cerevisiae
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