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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)
Publication Information
Food Science and Biotechnology / v.15, no.3, 2006 , pp. 453-457 More about this Journal
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
actinidine; activity; stability; kinetics; thermodynamics;
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Times Cited By KSCI : 2  (Citation Analysis)
Times Cited By Web Of Science : 2  (Related Records In Web of Science)
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1 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)   DOI
2 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)   DOI   ScienceOn
3 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)
4 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)   DOI
5 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)   DOI
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)   DOI
7 Mcdowell MA. Anionic proteinase from Actinidia Chinensis. Eur. J. Biochem. 14: 214-221 (1973)
8 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)   DOI
9 Arcus AC. Proteolytic enzyme of Actinidia Chinensis. Biochim. Biophys. Acta 33: 242-244 (1959)   DOI
10 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)   DOI
11 Lewis D, Luh BS. Development and distribution of actinidin in kiwifruit (Actinidia Chinensis) and its partial characterization. J. Food Biochem. 12: 109-116 (1988)   DOI
12 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)   DOI   ScienceOn
13 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)   DOI   ScienceOn
14 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)
15 Boland MJ, Hardman MJ. Kinetic studies on the thiol protease from Actinidia Chinensis. FEBS Lett. 27: 282-284 (1972)   DOI   ScienceOn
16 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)   DOI   ScienceOn
17 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)   DOI
18 Kim KY, Ustadi, Kim SM. Characteristics of the protease inhibitor purified from chum salmon (Oncorhynchus keta) eggs. Food Sci. Biotechnol. 15: 28-32 (2006)   과학기술학회마을
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)   DOI   ScienceOn
20 Lee OH, Kang SN, Lee BY. Rheological properties of dandelion root concentrations by extraction solvents. Food Sci. Biotechnol. 15: 33-38 (2006)   과학기술학회마을
21 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)   DOI   ScienceOn
22 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)   DOI   ScienceOn