DOI QR코드

DOI QR Code

Purification and Characterization of a Protease from Korean Pear (Pyrus serotina L.) as Meat Tenderizer

  • Guan, Hao-Li (Department of Food Science, Agricultural College of Yanbian University) ;
  • Mandal, P.K. (Department of Food Science and Biotechnology of Animal Resources, Konkuk University) ;
  • Lim, Hee-Kyong (Graduate School of Bio & Information Technology, Hankyong National University) ;
  • Baatartsogt, Oyungerel (Graduate School of Bio & Information Technology, Hankyong National University) ;
  • Lee, Chi-Ho (Department of Food Science and Biotechnology, of Animal Resources, Konkuk University) ;
  • Jeon, Gwang-Joo (Graduate School of Bio & Information Technology, Hankyong National University) ;
  • Choe, Il-Shin (School of Animal Life and Environment Science, Hankyong National University) ;
  • Choi, Kang-Duk (Graduate School of Bio & Information Technology, Hankyong National University)
  • Published : 2009.04.30

Abstract

This study was conducted for the isolation, purification, and characterization of a protease from Korean pear, to see its proteolytic activity on chicken actomyosin and to find the optimum pH and temperature of activity on chicken actomyosin. The protease was isolated from crude extract of Korean pear by ammonium sulfate precipitation. Further purification was done by DEAE-Sepharose ion-exchange chromatography, Mono-Q and Mini-Q column chromatography. The purified enzyme gave a single protein band on SDS polyacrylamide gel electrophoresis and the molecular weight was found to be 38 kDa. The specific activity of purified enzyme was 34,907 unit/mg with 25 fold purification and the yield was 2%. The purified enzyme incubated with chicken actomyosin showed high activity. The optimum pH and temperature for enzyme activity on chicken actomyosin were 6.5 and $70^{\circ}C$, respectively. A protease was purified from Korean pear for the first time and characterized. It was found to be promising for meat tenderization.

Keywords

References

  1. Blanchard, P. J. and Mantle, D. (1996) Comparison of proteolytic enzyme levels in chicken, pig, lamb and rabbit muscle at point of slaughter: role in meat tenderization post mortem. J. Sci. Food Agr. 71, 83-91 https://doi.org/10.1002/(SICI)1097-0010(199605)71:1<83::AID-JSFA552>3.0.CO;2-F
  2. Bradford, M. M. (1976) A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein dye-binding. Anal. Biochem. 72, 248- 254 https://doi.org/10.1016/0003-2697(76)90527-3
  3. Brooks, B. A., Kalsing, K. C. and Regenstein, J. M. (1985) Effects of antemortem injected crude papain in chicken muscle. J. Food Sc. 50, 1370-1375 https://doi.org/10.1111/j.1365-2621.1985.tb10479.x
  4. Choe, I. S. and Park, Y. J. (1996) A Study on the Utilization as Meat Tenderizer from Korean Pear Protease. Korean J. Food Sci. Ani. Resour. 16, 89-93
  5. Choe, I. S., Park, Y. J., Ishiroshi, M. and Samejima, K. (1996) A new protease in Korean pear as meat tenderizer. Animal Sci. Technol. 67, 443-446
  6. Gornall, A. G., Bardawill, J. C. and David, M. M. (1948) Determination of serum proteins by means of biuret reaction. J. Biol. Chem. 177, 751-758
  7. Han, S. K. and Chin, K. B. (2004) Study on meat tenderness of a protease extracted from domestic pear. Korean J. Food Sci. Ani. Resour. 24, 326-328
  8. Kang, C. K. and Rice, E. E. (1970) Degradation of various meat fractions by tenderizing enzymes. J. Food Sci. 35, 563 -567 https://doi.org/10.1111/j.1365-2621.1970.tb04809.x
  9. Kang, C. K. and Warner, W. D. (1974) Tenderization of meat with latex proteases. J. Food Sci. 39, 812-818 https://doi.org/10.1111/j.1365-2621.1974.tb17985.x
  10. Kee, H. J., Hwang, Y. S., Kim, K. H. and Hog, Y. H. (1998) Application fig protease to foods. Korean J. Food Sci. Ani. Resour. 18, 19-26
  11. Kim, E. M., Choe, I. S. and Hwang, S. G. (2003) Effects of singular or mixed type treatment of proteases isolated from Pear, Pineapple and Kiwifruit on actomyosin degradation. Korean J. Food Sci. Ani. Resour. 23, 193-199
  12. Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage, Nature 227, 680-685 https://doi.org/10.1038/227680a0
  13. Lee, D. H. and Jin, B. H. (2001) Encapsulation of bromelain in liposome. J. Food Sci. Nutr, 5, 81-85
  14. Lee, J. H. and Jang, H. R. (2005) Proteolytic properties of Sarcodon aspratus on Beef Loin. Korean J. Chulinal Res. 11, 1-14
  15. Marsh, B. B. and Carse, W. A. (1974) Meat tenderness and sliding-filament hypothesis. J. Food. Technol. 9, 129-139 https://doi.org/10.1111/j.1365-2621.1974.tb01756.x
  16. Miyada, D. S. and Tappel, A. L. (1956) The hydrolysis of beef proteins by various proteolytic enzymes. Food Res. 21, 217-223 https://doi.org/10.1111/j.1365-2621.1956.tb16913.x
  17. Nam, S. H., Walsh, M. K., and Yang, K. Y. (2006) The Enzymatic properties of Actinidine from Kiwifruit. Food Sci. Biotechnol. 15, 453-457
  18. Naveena, B. M. and Mendiratta, S. K. (2001) Tenderization of spent hen meat using ginger extract. Brit. Poultry Sci. 42, 344-350 https://doi.org/10.1080/00071660120055313
  19. Naveena, B. M, Mendiratta, S. K., and Anjaneyulu, A. S. R. (2004) Tenderization of buffalo meat using plant proteases from cucumis trigonus roxb(kachri) and zingerber offocinale roscoe (Ginger rhizome). Meat Sci. 68, 363-369 https://doi.org/10.1016/j.meatsci.2004.04.004
  20. Rampton, J. H., Pearson, A. M., Walker, J. E. and Kapsalis, J. G. (1971) Disc electrophoresis of Weber-Edsall extract and actomyosin from skeletal muscle. J. Agr. Food Chem. 19, 238-240 https://doi.org/10.1021/jf60174a046
  21. Roh, J. H., Kim, Y. B. and Kil, B. I. (2002) The Effect of Bulking Agent on Quality of Kiwifruit Powder in the Process of Domestic Kiwifruit Tenderizer. Korean J. Food Sci. Tech-nol. 34, 805-810
  22. Weber, K. and Osborn, M. (1969) The reliability of molecular weight determination by sodium dodecyl sulfate polyacrylamide gel electrophoresis, J. Biol. Chem. 244, 4406- 4412
  23. Yang, R., Kim, K. W., Lee, J. H., Youn, J. E., and Yu, J. H. (1975) Study on meat tenderizer part 1. Tenderizing ability of commercial meat tenderizer. Korean J. Food Sci. Technol. 7, 221-228

Cited by

  1. Functional characterization of purified pear protease and its proteolytic activities with casein and myofibrillar proteins vol.25, pp.S1, 2016, https://doi.org/10.1007/s10068-016-0095-6
  2. Isolation and characterization of a protease from the Actinidia arguta fruit for improving meat tenderness vol.25, pp.4, 2016, https://doi.org/10.1007/s10068-016-0171-y
  3. Identification and Functional Characterization of Cysteine Protease from Nine Pear Cultivars (Pyrus pyrifolia) vol.19, pp.7, 2016, https://doi.org/10.1080/10942912.2015.1107576
  4. The Physicochemical Characteristics of Marinated Beef Galbi under Different Cooking Conditions vol.28, pp.1, 2013, https://doi.org/10.7318/KJFC/2013.28.1.078
  5. Comparison of four purification methods to purify cysteine protease from Asian pear fruit (Pyrus pyrifolia) vol.5, 2016, https://doi.org/10.1016/j.bcab.2015.11.004