Food Science and Biotechnology

Korean Society of Food Science and Technology (한국식품과학회)
권호 표지

The Enzymatic Properties of Actinidine from Kiwifruit

  • Nam, Seung-Hee (School of Biological Science and Technology, Chonnam National University) ;
  • Walsh, Marie K. (Department of Nutrition and Food Sciences, Utah State University) ;
  • Yang, Kwang-Yeol (Department of Plant Biotechnology, College of Agriculture and Life Sciences, Chonnam National University)
  • Published : 2006.06.30
Download PDF
⟨ Previous Next ⟩

Abstract

Activity and stability of kiwifruit actinidine was determined in various conditions of pH, salt, and temperature using N-${\alpha}$-CBZ-lysine P-nitrophenyl ester as the substrate. Actinidine activity was low below pH 6, and undetectable below pH 3. The enzyme was stable in a pH range of 6.0-8.5. At $4^{\circ}C$ the enzyme was inactive in the presence of greater than 36% vinegar and in 2 M NaCl. Actinidine at $25^{\circ}C$ was unstable in 24% vinegar but stable in up to 3 M NaCl. With regard to freeze-thaw stability, actinidine retained 85% residual activity after being frozen at $-20^{\circ}C$ for 3 days. Based on Arrenius and Lineweaver-Burk plots, actinidine became unstable at greater than $45^{\circ}C$ with only 30% residual activity remaining after 6 min. The Km, kcat, and kcat/Km values of actinidine were $56\;{\mu}M$, 67/sec, and $1.2\;{\mu}M/sec$, respectively.

Keywords

References

  1. Arcus AC. Proteolytic enzyme of Actinidia Chinensis. Biochim. Biophys. Acta 33: 242-244 (1959) https://doi.org/10.1016/0006-3002(59)90522-0
  2. Boland MJ, Hardman MJ. Kinetic studies on the thiol protease from Actinidia Chinensis. FEBS Lett. 27: 282-284 (1972) https://doi.org/10.1016/0014-5793(72)80641-0
  3. Sugiyama S, Ohtsuki K, Kawabata M. Purification and characterization of six kiwifruit proteases isolated with two ion-exchange resins, Toyopearl-Super Q and Bakerbond WP-PEI. Biosci. Biotechnol. Biochem. 12: 1994-2000 (1996)
  4. Matulis D, Wu C, Pham TV, Guy C, Lovrien R. Protection of enzymes by aromatic sulfonates from inactivation by acid and elevated temperatures. J. Mol. Catal. B-Enzym. 7: 21-36 (1999) https://doi.org/10.1016/S1381-1177(99)00019-3
  5. Reid JD, Hussain S, Sreedharan SK, Bailey TS, Pinitglang S, Thomas EW, Verma CS, Brocklehurst K. Variation in aspects of cysteine proteinase catalytic mechanism deduced by spectroscopic observation of dithioester intermediates, kinetic analysis and molecular dynamics simulations. Biochem. J. 357: 343-352 (2001) https://doi.org/10.1042/0264-6021:3570343
  6. Kamphuis IG, Drenth J, Baker EN. Thiol proteases: comparative studies based on the high-resolution structures of papain and actinidine, and on amino acid sequence information for cathepsin B and H and stem bromelain. J. Mol. Biol. 182: 317-329 (1985) https://doi.org/10.1016/0022-2836(85)90348-1
  7. Mcdowell MA. Anionic proteinase from Actinidia Chinensis. Eur. J. Biochem. 14: 214-221 (1973)
  8. Pastorello EA, Conti A, Pravettoni V, Farioli L, Rivolta F, Ansaloni R, Ispano M, Incorvaia C, Giuffrida MG, Ortolani C. Identification of actinidine as the major allergen of kiwi fruit. J. Allergy Clin. Immunol. 101: 531-537 (1998) https://doi.org/10.1016/S0091-6749(98)70360-4
  9. Carne A, Moore CH. The amino acid sequence of the tryptic peptides from actinidine, a proteolytic enzyme from the fruit of Actinidia Chinensis. Biochem. J. 173: 83-83 (1978)
  10. Tello-Solis SR, Valle-Guadarrama MV, Hernandez-Arana A. Purification and circular dichroism studies of multiple forms of actinidine from Actinidia Chinensis (kiwifruit). Plant Sci. 106: 227-232 (1995) https://doi.org/10.1016/0168-9452(95)04085-9
  11. Salih E, Malthouse JPG, Kopwlessur S, Jarvis M, O'Driscoll M, Broklehurst K. Differences in the chemical and catalytic characteristics of two crystallographically identical enzyme catalytic sites. Biochem. J. 247: 181-193 (1987) https://doi.org/10.1042/bj2470181
  12. Lewis D, Luh BS. Development and distribution of actinidin in kiwifruit (Actinidia Chinensis) and its partial characterization. J. Food Biochem. 12: 109-116 (1988) https://doi.org/10.1111/j.1745-4514.1988.tb00363.x
  13. Patel M, Kayani IS, Mellor GW, Sreedharan S, Templeton W, Thomas EW, Thomas M, Brocklehurst K. Variation in the P2-S2 stereochemical selectivity towards the enantiomeric N-acetyl-phenylalanylglysine 4-nitronilides among the cysteine proteinases papin, ficin and actinidine. Biochem. J. 281: 553-559 (1992) https://doi.org/10.1042/bj2810553
  14. Topham CM, Salih E, Frazao C, Kowlessur D, Overington JP, Thomas M, Brocklehurst SM, Patel M, Thomas EW, Brocklehurst K. Structure-function relationships in the cysteine proteinases actinidine, papin and papaya proteinase ${\omega}$. Biochem. J. 280: 79-92 (1991) https://doi.org/10.1042/bj2800079
  15. Hussain S, Pinitglang S, Bailey TSF, Reid JD, Noble MA, Resminin M, Thomas EW, Greaves RB, Vermal CS, Brocklehurst K. Variation in the pH-dependent pre-steady-state and steady-state kinetic characteristics of cysteine-proteinase mechanism: evidence for electrostatic modulation of catalytic-site function by the neighbouring carboxylation anion. Biochem. J. 372: 735-746 (2003) https://doi.org/10.1042/BJ20030177
  16. Boland MJ, Hardman MJ. The actinidine-catalyzed hydrolysis of N-${\alpha}$-benzyloxycarbonyl-L-lysine-p-nitrophenyl ester: pH dependence and mechanism Eur. J. Biochem. 36: 575-582 (1973) https://doi.org/10.1111/j.1432-1033.1973.tb02947.x
  17. Anthon GE, Barrett DM. Kinetic parameters for the thermal inactivation of quality-related enzymes in carrots and potatoes. J. Agric. Food Chem. 50: 4119-4125 (2002) https://doi.org/10.1021/jf011698i
  18. Salih E, Broklehurst K. Investigation of the catalytic site of actinidine by using benzofuroxan as a reactive probe with selectivity for the thiolate-imidazolium ion-pair systems of cysteine proteinases. Biochem. J. 213: 713-718 (1983) https://doi.org/10.1042/bj2130713
  19. Sumner IG, Vaughan A, Eisenthal R, Pickersgill RW, Owen AJ, Goodenough PW. Kinetic analysis of papaya proteinase ${\omega}$. Biochim. Biophys. Acta 1164: 243-251 (1993) https://doi.org/10.1016/0167-4838(93)90255-P
  20. Brocklehurst K, Baines BS, Malthouse JPG. Differences in the interactions of the catalytic groups of the active centres of actinidine and papain. Biochem. J. 197: 739-746 (1981) https://doi.org/10.1042/bj1970739
  21. Kim KY, Ustadi, Kim SM. Characteristics of the protease inhibitor purified from chum salmon (Oncorhynchus keta) eggs. Food Sci. Biotechnol. 15: 28-32 (2006)
  22. Lee OH, Kang SN, Lee BY. Rheological properties of dandelion root concentrations by extraction solvents. Food Sci. Biotechnol. 15: 33-38 (2006)