Secretion and localization of Pseudomonas aurantiaca levansucrase expressed in Saccharomyces cerevisiae = Saccharomyces cerevisiae에서 발현된 Pseudomonas aurantiaca levansucrase의 분비국재성

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Title
Secretion and localization of Pseudomonas aurantiaca levansucrase expressed in Saccharomyces cerevisiae = Saccharomyces cerevisiae에서 발현된 Pseudomonas aurantiaca levansucrase의 분비국재성
Author(s)
C K Lim; K H Kim; Chul Ho Kim; Sang Ki Rhee; S W Nam
Bibliographic Citation
Korean Journal of (Applied) Microbiology & Biotechnology, vol. 32, no. 3, pp. 206-211
Publication Year
2004
Abstract
Levansucrase gene(lscA) from Pseudomonas aurantiaca was subcloned downstream of GAL1 promoter in pYES 2.0 and pYInu-AT [GAL10 promoter + exoinulinase signal sequence of Kluyveromyces marxianus], resulting pYES-lscA and pYInu-lscA, respectively. The two expression plasmids were introduced into an invertase-deficient strain, Sacchromayces cerevisiae SEY2102, and transformants with 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 8.62 U/ml with the strain harboring pYES-lscA and 5.43 U/ml with the strain harboring pYInu-lscA. The levansucrase activity of 80% was detected in the periplasmic space and cytoplasm. The levansucrase activity in the medium of SEY2102/pYInu-lscA was 0.87 U/ml whereas that of SEY2102/pYES-lscA was 0.47 U/ml, which implying the exoinulinase signal sequence didn't enhance the secretion efficiency of levansucrase. Furthermore, the recombinant levansucrase expressed in yeast seems to be produced as a hyper-glycosylated form.
Keyword
exoinulinase signalpseudomonas aurantiacasacchromayces cerevisiaelevansucrase
ISSN
0257-2389
Publisher
Korea Soc-Assoc-Inst
Type
Article
Appears in Collections:
Jeonbuk Branch Institute > Microbial Biotechnology Research Center > 1. Journal Articles
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