Browse > Article

Secretion and Localization of Pseudomonas auratiaca Levansucrase Expressed in Saccharomyces cerevisiae  

임채권 (동의대학교 미생물학과)
김광현 (동의대학교 미생물학과)
김철호 (한국생명공학연구원)
이상기 (한국생명공학연구원)
남수완 (동의대학교 생명공학과)
Publication Information
Microbiology and Biotechnology Letters / v.32, no.3, 2004 , pp. 206-211 More about this Journal
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 p YInu-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 transform ants 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.
Keywords
Pseudomonas aurantiaca; levansucrase; Sacchromayces cerevisiae; exoinulinase signal;
Citations & Related Records
Times Cited By KSCI : 4  (Citation Analysis)
Times Cited By SCOPUS : 0
연도 인용수 순위
1 Levansucrase of Rahnella aquatilis; cloning, expression, and levan formation /
[ Song, K. B.;J. W. Seo;M. G. Kim;S. K. Rhee ] / Ann. N. Y. Acad. Sci.   DOI
2 Overexpression of thermoal-kalophilic lipase from Bacillus stearothermophilus L1 in Saccharomyces cerevisiae /
[ Ahn, J. O.;H. W. Jang;H. W. Lee;E. S. Choi;S. J. Haam;T. K. Oh;J. K. Jung ] / J. Microbiol. Biotechnol.
3 Highly-efficient secretion of heterologous proteins from Saccharomyces cerevisiae using inulinase signal peptide /
[ Chung, B. H.;S. W. Nam;B. M. Kim;Y. H. Park ] / Biotechnol. Bioeng.   DOI
4 An MF<TEX>$\'{a}$</TEX>I-SUC2 (-factor-invertase) gene fusion for study of protein iocalization and gene expression in yeast /
[ Emr, S. D.;R. Schekman;M. C. Flessel;J. Thorner ] / Proc. Nati. Acad. Sci. USA   DOI   ScienceOn
5 Microbial levan /
[ Han, Y. W ] / Adv. Appl. Microbiol.   DOI
6 Comparison of characteristics of levan produced by different preparations of levansucrase from Zymomonas mobilis /
[ Jang, K. H.;K. B. Song;C. H. Kim;B. H. Chung;S. A. Kang;U. H. Chun;R. W. Choue;S. K. Rhee ] / Biotechnol. Lett.   DOI   ScienceOn
7 Molecular characterization of the levansucrase gene from Pseudomonas aurantiaca S-4380 and its expression in Escherichia coli /
[ Jang, E. K.;K. H. Jang;I. KOH;I. H. Kim;S. H. Kim;S. A. Kang;C. H. Kim;S. D. Ha;S. K. Rhee ] / J. Microbiol. Biotechnol.
8 Refolding and purification of Zymomonas mobilis levansucrase produced as inclusion bodies in fed-batch culture of recombinant Escherichia coli /
[ Kandlula, S.;B, H. Chung;K. H. Jang.;K. B. Song,C. H. Kim;S. K. Rhee ] / Protein Expression Purif.   DOI   ScienceOn
9 Cleavage of structural proteins during the assembly of the head of bacteriophage T4 /
[ Laemmli, U. K. ] / Nature   DOI   ScienceOn
10 Expression and secretion of Zymomonas mobilis levansucrase in Saccharomyces cerevisiae /
[ Lim, C. K.;Y. K. Kim;K. H. Kim;C. H. Kim;S. K. Rhee;S. W. Nam ] / J. Life Sci.   DOI
11 Use of dinitrosalicylic acid reagent for determination of reducing sugar /
[ Miller, G. L. ] / Anal. Chem.
12 Tansformation of intact yeast cells treated with alkali cations /
[ Ito, H.;Y. Fukuda;K. Murata;A. Kimura ] / J. Bacteriol.
13 Secretion and localization of invertase and inulinase in recombinant Saccharomyces cerevisiae /
[ Nam, S. W.;K. Yoda;M. Yamasaki ] / Biotechnol. Lett.   DOI   ScienceOn
14 Expression and localization of inulinase in recombinant Saccharomyces cerevisiae /
[ Nam, S. W.;M. H. Woo;B. M. Kim;B. H. Chung;Y. H. Park ] / Kor. J. Appl. Microbiol. Biotechnol.
15 Cloning, nucleotide sequence and expression in Escherichia coli of levansucrase genes from the plant pathogens Pseudomonas syringe pv. glycinea and P. syringe pv. phaseolicola /
[ Hettwer, U.;F. R. Jaeckel;J. Boch;M. Meyer;K. Rudolph;M. S. Ullrich ] / Appl. Environ. Microbiol.
16 The targeting of Bacillus subtilis levansucrase in yeast is correlated to both the hydrophobicity of the signal peptide and the net charge of the N-terminus mature part /
[ Scotti, P. A.;M. Praestegaard;R. Chambert;M. F. Petit-Glatron ] / Yeast   DOI   ScienceOn
17 Enzymatic synthesis of levan by Zymomonas mobilis levansucrase overexpressed in Escherichia coli /
[ Song, K. B.;S. K. Rhee ] / Biotechnol. Lett.
18 Xylan hydrolysis by treatment with endoxylanase and <TEX>$\beta$</TEX>-xylosidase expressed in yeast /
[ Heo, S. Y.;J. K. Kim;Y. M. Kim;S. W. Nam ] / J. Microbiol. Biotechnol.
19 Cloning and sequencing of the levansucrase gene from Acetobacter xylinum NCI 1005 /
[ Tajima, K.;T.Tanio;Y. Kobayashi;H. Kohno;M. Fujiwara;T. Shiba;T. Erata;M. Munekata;M. Takai ] / DNA Res.   DOI   ScienceOn
20 High-level secretion of human <TEX>$\'{a}_1$</TEX>-antitrypsin from Saccharomyces cerevisiae using inulinase signal sequence /
[ Kang, H. A.;S. W. Nam;K. S. Kwon;B. H. Chung;M. H. Yu ] / J. Biotechnol.   DOI
21 Expression and secretion of human serum abumin in the yeast Saccharomyces cerevisiae /
[ Kang, H. A.;M. S. Jung;W. K. Hong;J. H. Sohn;E. S. Choi;S. K. Rhee ] / J. Microbiol. Biotechnol.
22 Effect of glycine supplement on extracellular secretion of levansucrase from Pseudomonas aurantiaca S-4380 in recombinant Escherichia coli /
[ Kim, S. H.;E. K. Jang;I. H. Kim;K. H. Jang;S. A. Kang;B. I. Chang ] / Korean J. Biotechnol. Bioeng.
23 Characterization of levan produced by Serratia sp /
[ Kogima, I.;T. Saito;M. Iizuka;N. Minamiura;A. Ono ] / J. Ferment. Bioeng.   DOI   ScienceOn
24 Expression and secretion of recombinant inulinant inulinase under the control of GAL or GAP promoter in Saccharomyces cerevisiae /
[ Nam, S. W.;H. J. Lim;B. H. Chung;Y. K. Chang ] / Kor. J. Biotechnol. Bioeng.
25 Expression of Bacillus macerans cyclodextrin glucanotransferase gene in Saccharomyces cerevisiae /
[ Nam, S. W.;H. Y. Park;J. H. Seo;J. H. Kim;N. S. Han;B. W. Kim ] / Biotechnol. Lett.   DOI   ScienceOn
26 Characterization of Bacillus stearothermophilus cyclodextrin gluanotransferase that expressed by Saccharomyces cerevisiae /
[ Park, H. Y.;S. H. Jeon;H. Y. Kwon;S. W. Nam;H. W. Kim;K. H. Kim;B. W. Kim ] / Kor. J. Microbiol. Biotechnol.
27 Extracellular levansucrase of Bacillus subtilis produced in yeast remains in the cell under its precursor form /
[ Scotti, P. A.;R. Chambert;M. F. Petit-Glatron ] / Yeast   DOI   ScienceOn