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Further characterization and kinetic parameter determination of a milk-clotting protease fromMucor bacilliformis

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Abstract

Further characterization of an aspartyl protease fromMucor bacilliformis with milk-clotting activity was performed. An extinction coefficient, ε278 cm = 1.61 mL/mg/cm, a molecular mass of 35,400 Da and a pI of 5.2 were determined. Proteolytic activity and kinetic parameters were evaluated by using the hexapeptide Leu-Ser-pNO2-Phe-Nle-Ala-Leu-OMe as the substrate. The effect of pH and temperature on peptide cleavage, as well as protease heat stability, was determined. Such properties, taken as a whole, indicate that theM. bacilliformis protease can be considered a potential substitute for bovine chymosin in cheese manufacture.

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References

  1. Areces, L., Biscoglio de Jim’enez Bonino, M. J., Parry, M., Fraile, E. R., Fern’andez, H. M., and Cascone, O. (1992),Appl. Biochem. and Biotechnol. 37, 283–294.

    CAS  Google Scholar 

  2. Fern’andez Lahore, H. M., Miranda, M. V., Fraile, E. R., Biscoglio, M. J., and Cascone, O. (1995), ProcessBiochem. 30, 615–621.

    Article  Google Scholar 

  3. Martin, P., Raymond, M. N., Bricas, E., and Ribadeau Dumas, B. (1980),Biochim. Biophys. Acta 612, 410–420.

    CAS  Google Scholar 

  4. Sternberg, M. Z. (1971),J. Dairy Sci. 54, 159–167.

    CAS  Google Scholar 

  5. Lowry, O. H., Rosembrough, N. J., Farr, A. L., and Randall, R. J. (1951),J. Biol. Chem. 193, 265–275.

    CAS  Google Scholar 

  6. Arima, K., Yu, J., and Iwasaki, S. (1970), inMethods in Enzymology, vol. 19, Perlman, G. E. and Lorand L. eds., Academic, New York, pp. 446–459.

    Google Scholar 

  7. Laemmli, U. K. (1970),Nature 227, 680–685.

    Article  CAS  Google Scholar 

  8. Raymond, N. M., Bricas, E., Salesse, R., Garnier, J., Garnot, P., and Ribadeau Dumas, B. (1973),J. Dairy Sci. 56, 419–422.

    CAS  Google Scholar 

  9. Salesse, R. and Garnier, J. (1976),J. Dairy Sci. 59, 1215–1221.

    Article  CAS  Google Scholar 

  10. Umezawa, H. and Aoyagi, T. (1983), inProteinase Inhibitors: Medical and Biological Aspects, Katunuma, N., Umezawa, H., and Holze, H., eds., Springer-Verlag, Berlin, pp. 3–26.

    Google Scholar 

  11. Fruton, J. S. (1987), inHydrolytic Enzymes, Neuberger, A. and Brocklehnst, K., eds., Elsevier, Amsterdam, pp. 1–37.

    Google Scholar 

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Authors and Affiliations

  1. Instituto de Qu’imica y Fisicoqu’imica Biol’ogicas (UBA-CONICET), Facultad de Farmac’a y Bioqu’im’ica Junin 956, 1113, Buenos Aires, Argentina

    Graciela D. Venera, Claudia Machalinski, Hugo Zum’arraga, Mirtha J. Biscoglio & Jim’enez de Bonino

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  1. Graciela D. Venera
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  2. Claudia Machalinski
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  3. Hugo Zum’arraga
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  4. Mirtha J. Biscoglio
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  5. Jim’enez de Bonino
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Venera, G.D., Machalinski, C., Zum’arraga, H. et al. Further characterization and kinetic parameter determination of a milk-clotting protease fromMucor bacilliformis . Appl Biochem Biotechnol 68, 207–216 (1997). https://doi.org/10.1007/BF02785991

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  • DOI: https://doi.org/10.1007/BF02785991

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