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KMID : 0613820040140030429
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Journal of Life Science
2004 Volume.14 No. 3 p.429 ~ p.434
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Expression and Secretion of Zymomonas mobilis Levansucrase in Saccharomyces cerevisiae
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Lim Chae-Kwon
Kim Yi-Kyung
Kim Kwang-Hyeon
Kim Cheol-Ho
Lee Sang-Ki
Nam Soo-Wan
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Abstract
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Levansucrase gene (levU) from Zymomonas mobilis was subcloned downstream of GAL1 promoter in pYES 2.0 and pYInu-AT [GAL10 promoter＋exoinulinase signal sequence of Kluyveromyces marxianus], resulting pYES-levU and pYInu-levU, respectively. The two expression plasmids were introduced into an invertase-deficient strain, Saccharomyces cerevisiae SEY2102, and then transformants showing high activity of levansucrase were selected. When each yeast transformants was cultivated in medium containing galactose, the extracellular and intracellular activities of levansucrase reached about 7.17 U/ml with the strain harboring pYES-levU and 6.61 U/ml with the strain harboring pYInu-levU. It was found that about 50% of levansucrase were detected in the medium and periplasmic space, and exoinulinase signal sequence didn’t enhance the secretion efficiency. Furthermore, the recombinant levansucrase expressed in yeast seems to be produced as a hyper-glycosylated form.
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KEYWORD
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Zymomonas mobilis, levansucrase, Saccharomyces cerevisiae, exoinulinase signal
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