Expression, purification and characterization of porcine pancreatic Carboxypeptidase B from Pichia pastoris for the conversion of recombinant human insulin

Enzyme and Microbial Technology

Volumn 40, Issue 3, 2007, Pages 476-480

  Mansur, Manuel ^a   Martínez, Liliam ^a   Pérez, Mariela ^a   Alonso del Rivero, Maday ^b   Márquez, Isvaloy ^a   Proenza, Yanay ^a   Varas, Laura ^a   Avilés, Francesc X ^c  

^a CENTER FOR GENETIC ENGINEERING AND BIOTECHNOLOGY   (Cuba)

^b UNIVERSITY OF HAVANA   (Cuba)

^c UNIVERSITAT AUTÒNOMA DE BARCELONA   (Spain)

Author keywords

Carboxypeptidase B; Pichia pastoris; Protein secretion

Indexed keywords

AMINO ACIDS; INSULIN; ION EXCHANGE; PROTEINS; PURIFICATION; VECTORS;

CARBOXYPEPTIDASE B; CULTURE SUPERNATANT; PICHIA PASTORIS; PROTEIN SECRETION;

ENZYME KINETICS;

AMMONIUM ACETATE; ARGININE; CARBOXYPEPTIDASE B; COMPLEMENTARY DNA; ENZYME; FUNGAL ENZYME; LYSINE; MATING HORMONE ALPHA FACTOR; METHANOL; PANCREAS ENZYME; PEPTIDE; PREPROINSULIN; PROTEIN; RECOMBINANT HUMAN INSULIN; SEPHAROSE; TRYPSIN;

ARTICLE; CARBOXY TERMINAL SEQUENCE; ENZYME ANALYSIS; ENZYME KINETICS; ENZYME MECHANISM; ENZYME PURIFICATION; ENZYME RELEASE; ENZYME STABILITY; FUNGUS GROWTH; GENE VECTOR; GROWTH PERIOD; HORMONE RELEASE; ION EXCHANGE; NONHUMAN; PH; PICHIA PASTORIS; PROMOTER REGION; PROTEIN EXPRESSION; PROTEIN SECRETION; PURIFICATION; SACCHAROMYCES CEREVISIAE; SUPERNATANT; WILD TYPE; YEAST;

PICHIA PASTORIS; SACCHAROMYCES CEREVISIAE; SUS;

EID: 33846261892     PISSN: 01410229     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.enzmictec.2006.07.024     Document Type: Article

References (26)

1. Skidgel R.A. Structure and function of mammalian zinc carboxypeptidases. Zinc metalloproteases in health and disease (1996), Taylor and Francis, London p. 241-83
2. Folk J.E., and Gladner J.A. Carboxypeptidase B. I. Purification of the zymogen and specificity of the enzyme. J Biol Chem 231 (1958) 379-391
3. Tan A.K., and Eaton D.L. Activation and characterization of pro-Carboxypeptidase B from human plasma. Biochemistry 34 (1995) 5811-5816
4. Vendrell J., Avilés F.X., Fricker L.D., and Metallocarboxypeptidases. Handbook of metalloproteins vol. 3 (2004), John Wiley and Sons Ltd., Chichester p. 176-89
5. Villegas V., Vendrell J., and Avilés F.X. The activation pathway of Carboxypeptidase B from porcine pancreas: participation of the active enzyme in the proteolytic processing. Protein Sci 4 (1995) 1792-1800
6. Burgos F.J., Salva M., Villegas V., Soriano F., Méndez E., and Avilés F.X. Analysis of the activation process of porcine pro-Carboxypeptidase B and determination of the sequence of its activation segment. Biochemistry 30 (1991) 4082-4089
7. Plummer T.H., and Ryan T.J. A potent mercapto bi-product analogue inhibitor for human carboxypeptidase N. Biochem Biophys Res Commun 98 (1981) 448-454
8. Hass G.M., and Ryan C.A. Carboxypeptidase inhibitor from potatoes. Methods Enzymol 80 (1981) 778-791
9. Vendrell J., Querol E., and Avilés F.X. Metallocarboxypeptidases and their protein inhibitors. Structure, function and biomedical properties. Biochim Biophys Acta 1477 (2000) 284-298
10. Venhudova G., Canals F., Querol E., and Avilés F.X. Mutations in the N- and C-terminal tails of potato carboxypeptidase inhibitor influence its oxidative refolding process at the reshuffling stage. J Biol Chem 276 (2001) 11683-11690
11. Chance R.E., Kroeff E.P., Hoffmann J.A., and Frank B.H. Chemical, physical, and biologic properties of biosynthetic human insulin. Diabetes Care 4 (1981) 147-154
12. Thim L., Hansen M.T., Norris K., Hoegh I., Boel E., Forstrom J., et al. Secretion and processing of insulin precursors in yeast. Proc Natl Acad Sci USA 83 (1986) 6766-6770
13. Datar R., and Rosén C.G. Process economics downstream separation processes in biotechnology (1990), Marcel Dekker Inc., New York p. 741-93
14. Habermann P. Preparation of pancreatic pro-Carboxipeptidase B, isoforms and muteins thereof and their use. US Patent 6,531,294, 2003.
15. Folk J.E., and Gladner J.A. Carboxypeptidase B. II. Mode of action on protein substrates and its application of carboxyl terminal group analysis. J Biol Chem 231 (1958) 393-401
16. Ventura S., Villegas V., Sterner J., Larson J., Vendrell J., Hershberger C.L., et al. Mapping the pro-region of Carboxypeptidase B by protein engineering. Cloning, overexpression, and mutagenesis of the porcine proenzyme. J Biol Chem 274 (1999) 19925-19933
17. Cereghino L.J., and Cregg J.M. Heterologous protein expression in the methylotrophic yeast Pichia pastoris. FEMS Microbiol Rev 24 (2000) 45-66
18. Macauley-Patrick S., Fazenda L., McNeil B., and Harvey L.M. Heterologous protein production using the Pichia pastoris expression system. Yeast 22 (2005) 249-270
19. Tschopp J.F., Sverlow G., Kosson R., Craig W., and Grinna L. High level secretion of glycosylated invertase in the methylotrophic yeast Pichia pastoris. Biotechnology 5 (1987) 1305-1308
20. Paifer E., Margolles E., Cremata J., Montesino R., Herrera L., and Delgado J.M. Efficient expression and secretion of recombinant alpha amylase in Pichia pastoris using two different signal sequences. Yeast 10 (1994) 1415-1419
21. Rodríguez E., Mullaney E., and Le X. Expression of the Aspergillus fumigatus phytase gene in Pichia pastoris and characterization of the recombinant enzyme. Biochem Biophys Res Commun 268 (2000) 324-331
22. Becker W.M., and Guarente L. High-efficiency transformation of yeast by electroporation. Methods Enzymol 194 (1991) 182-187
23. Laemmli U.K. Cleavage of structural protein during the assembly of the head of bacteriophage T4. Nature 227 (1970) 680-685
24. Williams J., and Morrison J.F. The kinetics of reversible tight-binding inhibition. Methods Enzymol 63 (1979) 437-467
25. Boehm T., Pirie-Shepherd S., Trinh L.B., Shiloach J., and Folkman J. Disruption of the KEX1 gene in Pichia pastoris allows expression of full-length murine and human endostatin. Yeast 15 (1999) 563-572
26. Wolff E.C., Schirmer E.W., and Folk J.E. The kinetics of Carboxypeptidase B activity. J Biol Chem 237 (1962) 3094-3099