Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지

Expression of Carboxypeptidase Z cDNA from Absidia zychae in saccharomyces cerevisiae and its characteristics

Saccharomyces cerevisiae에서 Absidia zychae 의 Carboxypeptidase A cDNA 의 발현과 특성

  • 이병로 (우석대학교 식품공학과) ;
  • 김종화 (동경농공대학교 응용생물과학과)
  • Published : 1995.05.01
Download PDF
⟨ Previous Next ⟩

Abstract

Carboxypeptidase Z(CPZ) cDNA of Absidia zychae was experssed in Saccharomyces cerevisiae. The expressed CPZ(YCPZ) was secreted about 30 mg/l into the medium and has a little higher molecular weight than the wild type CPZ in SDS-PAGE. By the result of N-terminal amino acid sequencing, YCPZ has additional 15 amino acids residues in N-terminus of CPZ. But YCPZ shows no difference with CPZ in enzyme activity and substrate specificity. For the identification of processing mechanism of YCPZ, 36-Arg was changed to 36-Thr by site specific mutagenesis. Mutant YCPZ does not processed at 36-Thr. It was, therefore, concluded that the YCPZ was processed by KEX2. According to endo F treatment, high amount of carbohydrate was N-glycosylated in YCPZ.

Keywords

References

  1. Carlsberg Res. Commun. v.51 Serine carboxypeptidases. A review Breddam,K.
  2. Cell v.48 Protein sorting in yeast: The localization determinant of yeast vacuolar carboxypeptidase Y resides in the propeptide Valls,L.A.;Hunter,C.P.;Rothman,J.H.;Stevens,T.H.
  3. Carlsberg Res. Commun. v.51 Primary sructure of carboxypeptidase I from malted barley Sorensen,S.B.;Breddanr,K.;Svendsen,I.
  4. Carlsberg Res. Commun. v.52 Primary structure of carboxypeptidase Ⅱ from malted barley Sorensen,S.B.;Svendsen,I.;Breddanr,K.
  5. Carlsberg Res. Commun. v.54 Primary structure of carboxypeptidase Ⅲ from malted barley Sorensen,S.B.;Svendsen,I.;Breddanr,K.
  6. Carlsberg Res. Commun. v.52 Primary structure and enzymatic properties of carboxypeptidase Ⅱ from wheat bran Breddanr,K.;Sorensen,S.B.;Svendsen,I.
  7. J. Biol. Chem. v.262 A gibberellin-responsive wheat gene has homology to yeast carboxypeptidase Y. Baulcombe,D.C.;Barker,R.F.;Jarvis,M.G.
  8. Plant Mol. Biol. v.19 Structure and expression during the germination of rice seeds of the gene for a carboxypeptidase Washio,K.;Ishikawa,K.
  9. Curr. Genet. Molecular cloning and sequence analysis of the scpZ gene encoding serine carboxypeptidase of Absidia zychae Lee,B.R.;Takeuchi,M.;Kobayashi,Y.
  10. J. Biol. Chem. v.265 Structure of wheat serin carboxypeptidase Ⅱ at 3.5-Å resolution Liao,D.I.;Remington,S.J.
  11. Biochemistry v.31 Refined atomic model of wheat serine carboxypeptidase Ⅱ at 2.2-Å resolution Liao,D.I.;Breddam,K.;Sweet,R.M.;Bullock,T.;Remington,S.T.
  12. Biosci. Biotech. Biochem. v.57 Purification and characterization of serine carboxypeptidases from Absidia zychae Lee,B.R.;Takeuchi,M.;Kobayashi,Y.
  13. Genetics v.122 A system of shuttle vectors and yeast host strain designed for efficient manipulation of DNA in Saccharomyces cerevisiae Sikorski,R.S.;Hieter,P.
  14. Methods in Enzymology v.194 Vectors for constitutive and inducible gene expression in yeast Schena,M.;Picard,D.;Yamamoto,K.R.
  15. J. Bacteriol. v.153 Transformation of intact yeast cells treated with alkali cations Ito,H.;Fukuda,Y.;Murata,K.;Kimura,A.
  16. Proc. Natl. Acad. Sci. U.S.A. v.76 Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications Towbin,H.;Staehelin,T.;Gordon,J.
  17. Proc. Natl. Acad. Sci. U.S.A. v.82 Rapid and efficient site-specific mutagenesis without phenotypic selection Kunkel,T.A.
  18. J. Biol. Chem. v.262 Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes Matsudaira,P.
  19. Science v.239 Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase Saiki,R.K.;Gelfand,D.H.;Stoffer,S.;Scharf,S.J.;Higuchi,R.;Horn,G.T.;Mullis,K.B.;Erlich,H.A.
  20. Korean J. Biotechnol. Bioeng. v.8 Multiple forms of serine-type carboxypeptidase produced by Absidia zychae Lee,B.R.;Ahn,B.Y.
  21. Science v.241 Yeast KEX2 endopeptidase correctly cleaves a neuroendocrine prohormone in mammalian cells Thomas,G.;Thorne,B.A.;Thomas,L.;Allen,R.G.;Hruby,D.E.;Fuller,R.;Thorner,J.
  22. Biochim. Biophys. Acta. v.258 Purification and characterization of a new type of acid caboxypeptidase from Aspergillus Ichishima,E.