한국미생물·생명공학회지 (Microbiology and Biotechnology Letters)

  • 제25권3호
  • /
  • Pages.258-265
  • /
  • 1997
  • /
  • 1598-642X(pISSN)
  • /
  • 2234-7305(eISSN)
한국미생물·생명공학회 (The Korean Society for Microbiology and Biotechnology)
권호 표지

재조합 Saccharomyces cerevisiae에서 Inulinase와 Invertase의 발현과 분비에 미치는 배양조건의 영향

  • 남수완 (동의대학교 미생물학과) ;
  • 신동하 (동의대학교 미생물학과) ;
  • 김연희 (동의대학교 미생물학과)
  • 발행 : 1997.06.01
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

The effects of medium pH and culture temperature on the expression and secretion of inulinase and invertase were investigated with recombinant Saccharomyces cerevisiae cells. These cells were obtained by transformation of 2$\mu$-based plasmids pYI10 and pYS10 which contain Kluyveromyces marxianus inulinase gene (INU1A) and S. cerevisiae invertase gene (SUC2), respectively, in the downstream of GAL1 promoter. The expression level and localization of inulinase and invertase were not affected significantly by the initial medium pH: secretion efficiencies of inulinase and invertase into the medium were about 90% and 60%, respectively, in the pH ranges of 4.0 to 6.5. However, the expression and secretion of both enzymes were strongly dependent on the culture temperature. The highest expression (7.7 units/mL) and secretion (6.7 units/mL) of inulinase were observed at 28$\circ$C and 30$\circ$C. As a consequence of decreased localization of inulinase in the periplasmic space, the secretion efficiency increased from 68% at 20$\circ$C, to 95% at 35$\circ$C,. The total expression level and secretion efficiency of invertase increased from 19 units/mL and 55% at 20$\circ$C to 25 units/mL and 68% at 35$\circ$C, respectively. Irrespective of the culture temperature, the invertase activity in the cellular fraction (periplasmic space and cytoplasmic fractions) was kept constant at around 33-45%.

키워드

참고문헌

  1. Yeast. v.7 Passage of molecules through yeast cell walls: a brief essay-review De Nobel, J. G.;J. A. Barnett
  2. Yeast. v.9 In Saccharomyces cerevisiae, protein secretion into the growth medium depends on environmental factors Rossini, D.;D. Porro;L. Brambilla;M. Venturini;B. M. Ranzi;M. Vanoni;L. Alberghina
  3. Proc. Natl. Acad. Sci. USA v.81 Regulated expression of a human interferon gene in yeast: control by phosphate concentration or temperature Kramer, R. A.;T. M. DeChiara;M. D. Schaber;S. Hilliker
  4. Biotechnol. Bioeng. v.29 Controlled expression and purification of human immune interferon from high-cell-density fermentations of Saccharomyces cerevisiae Fieschko, J. C.;K. M. Egan;T. Ritch;R. A. Koski;M. Jenes;G. A. Bitter
  5. Biotechnol. Bioeng. v.35 A process for the production of human proinsulin in Saccharomyces cerevisiae Tottrup, H. V.;S. Carlsen
  6. J. Biol. Chem. v.265 Characterization of a nonglycosylated single chain urinary plasminogen activator secreted from yeast Melnick, L. M.;B. G. Turner;P. Puma;B. Price-Tillotson;K. A. Salvato;D. R. Dumais;D. T. Moir;R. J. Broeze;G. C. Avgerinos
  7. Agri. Biol. Chem. v.53 The effects of culture conditions on the secretion of human lysozyme by Saccharomyces cerevisiae A2-1-1A Ichikawa, K.;K. Komiya;K. Suzuki;T. Nakahara;Y. Jigami
  8. Appl. Microbiol. Biotechnol. v.34 Expression of human blood coagulation factor XIIIa in Saccharomyces cerevisiae: dependence of the expression levels from host-vector systems and medium compositions Broker, M.;O. Bauml;A. Gottig;J. Ochs;M. Bodenbenner;E. Amann
  9. J. Microbiol. Biotechnol. v.3 Effect of galactose and dextrose on human lipocortin-1 expression in recombinant Saccharomyces cerevisiae carrying galactose-regulated expression system Nam, S. W.;D. J. Seo;S.-K. Rhee;Y. H. Park;B. H. Chung
  10. バイオサイエンスとインダストリ- v.49 酵母によるヒト福經成長因子の分泌生産 Fumio, H.
  11. Cell. v.28 Two differentialy regulated mRNAs with different 5' ends encode secreted and intracellular forms of yeast invertase Carlson, H.;D. Botstein
  12. J. Biol. Chem. v.259 Early step in processing of yeast glycoprotein Esmon, B.;P. C. Esmon;R. Schekman
  13. Ann. Rev. Cell. Biol. v.1 Protein localization and membrane traffic in yeast Schekman, R.
  14. J. Biol. Chem. v.262 Structure, assembly, and secretion of octameric invertase Esmon, P. C.;B. E. Esmon;I. E. Schauer;A. Taylor;R. Schekman
  15. Ph.D. Thesis, The University of Tokyo Studies on the extracellular secretion of proteins in yeast Ko, J. H.
  16. FEBS Lett. v.289 Cloning and sequencing of the inulinase gene of Kluyeromyces marxianus var. marxianus ATCC 12424 Laloux, O.;J. P. Cassart;J. Delcour;J. van Beeumen;J. Vantenhaute
  17. J. Microbiol. Serol. v.26 Studies on a β-fructosidase(inulinase) produced by Saccharomyces fragilis. Antonie van Leeuwenhoek Snyder, H. E.;H. J. Phaff
  18. Appl. Environ. Microbiol. v.54 Production, distribution, and kinetics properties of inulinase in continuous cultures of Kluyveromyces marcianus CBS 6556 Rouwenhorst, R. J.;L. E. Visser;A. A. van der Baan;W. A. Scheffers;J. P. van Dijken
  19. Appl. Environ. Microbiol. v.56 Structure and properties of the extracellular inulinase of Kluyveromyces marxianus CBS 6556 Rouwenhorst, R. J.;M. Hensing;J. Verbakel;L. E. Visser;W. A. Scheffers;J. P. van Dijken
  20. Proc. Natl. Acad. Sci. USA v.80 An MFα1-SUC2(α-factor-invertase) gene fusion for study of protein localization and gene expression in yeast Emr, S. D.;R. Schekman;M. C. Flessel;J. Thorner
  21. Biotechnol. Lett. v.15 Secretion and localization of invertase and inulinase in recombinant Saccharomyces cerevisiae Nam, S. W.;K. Yoda;M. Yamasaki
  22. J. Bacteriol. v.153 Transformation of intact yeast cells treated with alkali cations Ito, H.;Y. Fukuda;K. Murata;A. Kimura
  23. Anal. Chem. v.55 Use of dinitrosalicylic acid reagent for determination of reducing sugar Miller, G. L.
  24. Biotechnol. Bioeng. v.40 Fermentation kinetics of recombnant yeast in batch and fed-batch cultures Patkar, A.;J.-H. Seo
  25. Foods Biotechnol. v.4 Effects of environmental variables on expression and secretion of cloned invertase from recombinant Saccharomyces cerevisiae Marten, M. R.;C. M. Choi;J. H. Seo
  26. J. Bacteriol. v.173 Removal of N-glycosylation sites of yeast acid phosphatase severely affects protein folding Riederer, M. A.;A. Hinnen
  27. Appl. Environ. Microbiol. v.56 Localization of inulinase and invertase in Kluyveromyces species Rouwenhorst, R. J.;W. S. Ritmeester;W. A. Scheffers;J. P. van Dijken
  28. Kor. J. Appl. Microbiol. Biotechnol. v.22 Expression and localization of inulinase in recombinant Saccharomyces cerevisiae Nam, S. W.;M. H. Woo;B. M. Kim;B. H. Chung;Y. H. Park
  29. Kor. J. Biotechnol. Bioeng. v.11 Expression and secretion of recombinant inulinase under the control of GAl or GAP promoter in Saccharomyces cerevisiae Nam, S. W.;H. J. Lim;B. H. Chung;Y. K. Chang