Skip to main content

Isolation and characterization of a protease from the Actinidia arguta fruit for improving meat tenderness

Food Science and Biotechnology volume 25, pages 1059–1064 (2016)Cite this article

Abstract

An protease from Actinidia arguta for improving meat tenderness was purified, characterized from wild A. arguta fruit by ammonium sulfate precipitation, Sephdex G-25 gel filtration chromatography, and DEAE Sepharose Fast Flow ion exchange chromatography, and its activity was investigated. The purified protease was subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis to obtain a single band of protease. The protease was purified successfully, and found to have a molecular weight of 23.8 kDa (mass spectrometry). The specific activity of the purified protease reached 53,428 U/mg with a 25.5-fold purification factor and 9% activity recovery. Based on N-terminal sequencing results, the A. arguta protease was derived from the class of actinidia proteases that have an N-terminal sequence of VLPDY VDWRS AGAVV. The protease was effective for tenderizing beef and decomposing actomyosin, suggesting the potential application for improving meat tenderness.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

43,34 €

Price includes VAT (Finland)
Tax calculation will be finalised during checkout.

References

  1. Huffman KL, Miller MF, Hoover LC, Wu CK, Briton HC, Ramsey CB. Effect of beef tenderness on consumer satisfaction with steaks consumed in the home and restaurant. J. Anim. Sci. 74: 91–97 (1996)

    CAS  Article  Google Scholar 

  2. Marsh BB, Carse WA. Meat tenderness and sliding-filament hypothesis. Int. J. Food Sci. Tech. 9: 129–139 (1974)

    Article  Google Scholar 

  3. Savell JW, Cross HR. Animal product options in the market place. pp. 345–355. In: The role of fat in the palatability of beef, pork, and lamb. Council NR (ed). National Academy Press, Washington DC, USA (1988)

    Google Scholar 

  4. Brooks BA, Kalsing KC, Regenstein JM. Effects of antemortem injected crude papain in chicken muscle. J. Food Sci. 50: 1370–1375 (1985)

    CAS  Article  Google Scholar 

  5. Kee HJ, Hwang YS, Kim KH, Hog YH. Application fig protease to foods. Korean J. Food Sci. An. 18: 19–26 (1998)

    Google Scholar 

  6. Lee DH, Jin BH. Encapsulation of bromelain in liposome. J. Food Sci. Nutr. 5: 81–85 (2001)

    Google Scholar 

  7. Kim EM, Choe LS, Hwang SG. Effects of singular or mixed type treatment of proteases isolated from pear, pineapple and kiwifruit on actomyosin degradation. Korean J. Food Sci. An. 23: 193–199 (2003)

    Google Scholar 

  8. Rødbotten R, Solgaard K, Johannessen TC. Effect of kiwi fruit powder in tenderizing M. Triceps brachii from mature cows. Meat Sci. 101: 117 (2015)

    Google Scholar 

  9. Arcus AC. Proteolylic enzymes of Actinidia chinensis. Biochim. Biophys. Acta 33: 241–243 (1959)

    Article  Google Scholar 

  10. Bolank MJ, Herdnan MJ. Kinetic studies on the thiol protease from Actinidia chinensis. FEBS Lett. 27: 251–254 (1972)

    Article  Google Scholar 

  11. Carne A, Moore CH. The amino acid sequence of the tryptic peptides from Actinidin, a proteolytic enzyme from the fruit of Actinidia chinensis. J. Biochem. 173: 73–78 (1978)

    CAS  Article  Google Scholar 

  12. Brocklehurst K, Baines BS, Malthouse JPG. Differences in the interactions of the catalytic groups of the active centers of actinidin and papain. J. Biochem. 197: 739–741 (1981)

    CAS  Article  Google Scholar 

  13. Edward N, Baker MJ, Boland PC. The specificity of actinidin and its relationship to the structure of the enzyme. BBA -Enzymology 616: 30–34 (1980)

    Google Scholar 

  14. Ha M, Bekhit AA, Carne A, Hopkins DL. Characterisation of commercial papain, bromelain, actinidin and zingibain protease preparations and their activities toward meat proteins. Food Chem. 134: 95–105 (2012)

    CAS  Article  Google Scholar 

  15. Kim JY, Lee IK, Son MW. Effects of orally administered Actinidia arguta fruit extract on 2-chloro-1,3,5-trinitrobenzene-induced atopic dermatitis like skin lesions in NC/Nga mice. J. Med. Food 12: 1004–1015 (2009)

    CAS  Article  Google Scholar 

  16. Weber K, Osborn M. The reliability of molecular weight determination by sodium dodecyl sulfate polyacrylamide gel electrophoresis. J. Biol. Chem. 244: 4406–4412 (1969)

    CAS  Google Scholar 

  17. Rampton JH, Pearson AM, Walker JE, Kapsalis JG. Disc electrophoresis of Weber-Edsall extract and actomyosin from skeletal muscle. J. Agr. Food Chem. 19: 238–240 (1971)

    CAS  Article  Google Scholar 

  18. Li GH, Mandal PK, Lim H, Baatartsogt O, Chi HL, Jeon SC. Choi K. Purification and characterization of a protease from Korean pear (Pyrus serotina L) as meat tenderizer. Korean J. Food Sci. An. 29: 157–163 (2009)

    CAS  Article  Google Scholar 

  19. Kyung HC. Amino-acid sequence and glycan structures of cysteine proteases with proline specificity from ginger rhizome Zingiber off icinale. Eur. J. Biochem. 267: 1516–1526 (2000)

    Article  Google Scholar 

  20. Bradford MM. A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein dye-binding. Anal. Biochem. 72: 248–254 (1976)

    CAS  Article  Google Scholar 

  21. Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ. Gapped BLAST and PSIBLAST: A new generation of protein database search programs. Nucleic Acids Res. 25: 3389–3402 (1997)

    CAS  Article  Google Scholar 

  22. Miyada DS, Tappel AL. The hydrolysis of beef proteins by various proteolytic enzymes. Food Res. Int. 21: 217–223 (1956)

    CAS  Article  Google Scholar 

  23. Kang CK, Rice EE. Degradation of various meat fractions by tenderizing enzymes. J. Food Sci. 35: 563–567 (1970)

    CAS  Article  Google Scholar 

  24. Xiang D, Zhang J, Chen Y. Expressions and purification of a mature form of recombinant human chemerin in escherichia coli. Protein Expres. Purif. 69: 153–158 (2010)

    CAS  Article  Google Scholar 

  25. Nieuwenhuizen N, Beuning L, Sutherland P, Sharma N, Cooney J, Bieleski L, Schroeder R, MacRae E, Atkinson R. Identification and characterisation of acidic and novel basic forms of actinidin, the highly abundant cysteine protease from kiwifruit Funct. J. Plant Biol. 34: 946–961 (2007)

    CAS  Google Scholar 

Download references

Author information

Affiliations

  1. College of Agriculture, Yanbian University, Yanji, Jilin, 133002, China

    Juan Wang, Haoming Liu, Haili Wang, Mingxun Cui, Qing Jin, Tie Jin, Fushun Cui, Taihua Cui, Chengyun Liang & Guanhao Li

  2. Food Research Center, Yanbian University, Yanji, Jilin, 133002, China

    Juan Wang, Qing Jin, Tie Jin, Fushun Cui, Taihua Cui, Chengyun Liang & Guanhao Li

  3. Collaborative Innovation Center of Beef Cattle Science and Major Demand for Industrial Technology, Yanbian University, Yanji, Jilin, 133002, China

    Qing Jin, Fushun Cui, Chengyun Liang & Guanhao Li

  4. School of Food Science, Kyungil University, Gyeongsan, Gyeongbuk, 38428, Korea

    Bumsik Kim

Authors
  1. Juan Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Haoming Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Haili Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Mingxun Cui
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Qing Jin
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Tie Jin
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Fushun Cui
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Taihua Cui
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Chengyun Liang
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Bumsik Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Guanhao Li
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Guanhao Li.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Wang, J., Liu, H., Wang, H. et al. Isolation and characterization of a protease from the Actinidia arguta fruit for improving meat tenderness. Food Sci Biotechnol 25, 1059–1064 (2016). https://doi.org/10.1007/s10068-016-0171-y

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10068-016-0171-y

Keywords

  • plants
  • Actinidia arguta
  • protease
  • meat
  • tenderness