Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Purification, Characterization, and Inhibitory Activity of Glassfish (Liparis tanakai) Egg High Molecular Weight Protease Inhibitor Against Papain and Cathepsin

  • Ustadi Ustadi (Fisheries and Marine Department, Agriculture Faculty, Gadjah Mada University) ;
  • You Sang-Guan (Faculty of Marine Bioscience and Technology, Kangnung National University) ;
  • Kim Sang-Moo (Faculty of Marine Bioscience and Technology, Kangnung National University)
  • Published : 2006.04.01
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Abstract

Two protease inhibitors of 67 and 18 kDa, respectively, were purified from the eggs of glass fish, Liparis tanakai, by affinity chromatography and electro-elution method. The high molecular weight (HMW) protein was purified with a specific inhibitory activity, yield, and purity of 18.46 U/mg, 0.07%, and 131.86 fold, respectively, and was further characterized: Optimal temperature and pH for inhibitory activity of the HMW glassfish egg protease inhibitor were $40^{\circ}C$ and pH 6, respectively, and it was stable between $5^{\circ}C\;and\;50^{\circ}C$ in the pH range of 5-6 with maximal stability at pH 6. It was shown to be a competitive inhibitor against papain with an inhibition constant $(K_i)$ of 97.02 nM. Moreover, the 67 kDa protein inhibited cathepsin, a cysteine protease, more effectively than did an egg-white protease inhibitor. The HMW glassfish egg protease inhibitor was classified as a member of the family III (kininogen).

Keywords

References

  1. Abrahamson, M., A. Ritonja, M. A. Brown, A. Grubb, W. Machleidt, and A. J. Barrett. 1987. Identification of the probable inhibitory reactive sites for the cysteine protease inhibitor human cystatin C and chicken cystatin. J. Biol. Chem. 262: 9688-9694
  2. An, H., M. Y. Peters, and T. A. Seymour. 1996. Roles of endogenous enzymes in surimi gelation. J. Food Sci. 7: 321-326
  3. Barret, A. J., N. D. Rawlings, M. E. Davies, W. Macleidt, G.. Salvesen, and V. Turk. 1986. Cysteine protease inhibitors of the cystatin superfamily, pp. 515-569. In A. J. Barrett and D. Salvesen (eds.), Proteinase Inhibitors. Elsevier, Amesterdam
  4. Barrett, A. J. and H. Kirschke. 1981. Cathepsin B, Cathepsin H. and Cathepsin L, pp. 535-561. In A. J. Barrett (ed.), Methodes in Enzymology, vol. 80. Academic Press, San Diego, LA, U.S.A
  5. Borla, O. O., C. B. Martone, and J. J. Sanchez. 1998. Protease I inhibitor system in fish muscle: A comparative study. Comp. Biochem. Physiol. B 119: 101-105 https://doi.org/10.1016/S0305-0491(97)00282-4
  6. Brillard-Bourdet, M., V. Nguyen, M. Ferrer-Di Martino, F. Gauthierand, and T. Moreau. 1998. Purification and characterization of a new cystatin from Taiwan cobra (Naja naja atra) venom. Biochem. J. 331: 239-244 https://doi.org/10.1042/bj3310239
  7. Conlon, J. M. and K. R. Olson. 1993. Purification of a vasoactive peptide related to lysyl-bradykinin from trout plasma. FEBS Lett. 334: 75-78 https://doi.org/10.1016/0014-5793(93)81684-R
  8. Conlon, J. M., B. Platzack, L. E. Marra, J. H. Youson, and K. R. Olson. 1995. Isolation and biological activity of [Trp5]bradykinin from the plasma of the phylogenetically ancient fish, the bowfin and the longnosed gar. Peptides 16: 485-489 https://doi.org/10.1016/0196-9781(94)00202-H
  9. Dixon, M. and E. C. Webb. 1979. Enzymes. Logman Group Ltd., London, U.K
  10. Gruden, K., B. Strukelj, T. Popovie, B. Lenareie, T. Bevec, J. Brzin, I. Kregar, J. Herzog-Velikonja, W. J. Stiekema, D. Bosch, and M. A. Jongsma. 1998. The cysteine activity of Colorado potato beetle (Leptinotarsa decemlineata Say) guts, which is incentive to potato protease inhibitor, is inhibited by thyroglobulin type-1 domain inhibitors. Insect Biochem. Mol. Biol. 28: 549-560 https://doi.org/10.1016/S0965-1748(98)00051-4
  11. Hammann, D. D., P. M. Amato, M. C. Wu, and E. A. Foegeding. 1990. Inhibition of modori (gel weakening) in surimi by plasma hydrolysate and egg white. J. Food Sci. 55: 665-669 https://doi.org/10.1111/j.1365-2621.1990.tb05202.x
  12. Sri, R., F. Tanuwidjaja, Y. Rukayadi, A. Suwanto, M. T. Suhartono, J. K. Hwang, and Y. R. Pyun. 2004. Study of thermostable chitinase enzymes from indonesian Bacillus K29-14. J. Microbiol. Biotechnol. 14: 647-652
  13. Hernandez, A. A. and W. R. Roush. 2002. Recent advance in the synthesis, design and selection of cysteine protease inhibitors. Curr. Opin. Chem. Biol. 6: 459-465 https://doi.org/10.1016/S1367-5931(02)00345-9
  14. Jacinto, T., K. V. S. Fernandes, O. L. T. Machado, and C. L. Siqueira Jr. 1998. Leaves of transgenic tomato plants overexpressing prosystemin accumalate high levels of cystatin. Plant Sci. 138: 35-42 https://doi.org/10.1016/S0168-9452(98)00153-8
  15. Jiang, K., J. Lee, and H. Chen. 1994. Purification and characterization of cathepsin B from ordinary muscle of mackerel (Scomber australasicus). J. Agric. Food Chem. 42: 1073-1079 https://doi.org/10.1021/jf00041a005
  16. Jiang, S. T., J. J. Lee, and C. Y. Tsao. 1997. Mackerel cathepsin B and L effect on thermal degradation of surimi. J. Food Sci. 62: 1-6 https://doi.org/10.1111/j.1365-2621.1997.tb04356.x
  17. Laemmli, U. K. 1970. Cleavage of structural protein during the assembly of the head of bacteriophagety. Nature 227: 265-275 https://doi.org/10.1038/227265a0
  18. Lanier, T. C. 2000. Surimi gelation chemistry, pp. 237-266. In J. W. Park (ed.), Surimi and Surimi Seafood. Marcel Dekker Inc., New York, NY, U.S.A
  19. In, M.-J., E.-S. Jang, Y.-J. Kim, and N.-S. Oh. 2004. Purification and properties of an extracellular acid phytase from Pseudomonas fragi Y9451. J. Microbiol. Biotechnol. 14: 1004-1008
  20. Lenarcic, B., A. Ritonja, I. Dolenc, V. Stoka, S. Berbic, J. Pungercar, B. Strukelj, and V. Turk. 1993. Pig leukocyte cysteine proteinase inhibitor (PLCPI), a new member of the stefin family. FEBS Lett. 336: 289-292 https://doi.org/10.1016/0014-5793(93)80822-C
  21. Li, Z., S. M. Secor, V. A. Lance, M. A. Masini, M. Vallarino, and J. M. Conlon. 1998. Characterization of bradykininrelated peptides generated in the plasma of six sarcopterygian species (African lungfish, amphiuma, coachwhip, bullsnake, gila monster, and Gray's monitor). Gen. Comp. Endocrinol. 112: 108-114 https://doi.org/10.1006/gcen.1998.7149
  22. Michaelis, L. and M. L. Menten. 1913. Die Kinetik der invertinwerkung. Biochem. Z. 49: 333-369
  23. Olenen, A., N. Kalkkinen, and L. Paulin. 2003. A new type of cysteine proteinase inhibitor - the salrin gene from Atlantic salmon (Salmo salar L) and Arctic charr (Salvelinus alpinus). Biochemie 1: 001-005
  24. Otto, H. H. and T. Schirmeister. 1997. Cysteine proteases and their inhibitors. Chem. Rev. 97: 133-171 https://doi.org/10.1021/cr950025u
  25. Platzack, B. and J. M. Conlon. 1997. Purification, structural characterization, and cardiovascular activity of cod bradykinins. Am. J. Physiol. 272: R710-R717
  26. Jung, H. K. and S.-D. Kim. 2005. An antifungal antibiotic purified from Bacillus megaterium KL39, a biocontrol agent of red-pepper Phytophthora-blight disease. J. Microbiol. Biotechnol. 15: 1001-1010
  27. Ritonja, A., W. Machleidt, and A. J. Barrett. 1985. Amino acid sequence of the intracellular cysteine protease inhibitor cystatin B from human liver. Biochem. Biophys. Res. Commun. 131: 1187-1192 https://doi.org/10.1016/0006-291X(85)90216-5
  28. Ritonja, A., T. H. Coetzer, R. N. Pike, and C. Dennison. 1996. The amino acid sequence, structure comparisons and inhibition kinetics of cathepsin L and sheep stefin B. Comp. Biochem. Physiol. B Biochem. B. 114: 193-198 https://doi.org/10.1016/0305-0491(96)00010-7
  29. Sangorrin, M. P., E. J. Folco, C. M. Martone, and J. J. Sanchez. 2001. Purification and characterization of a protease inhibitor from white croaker skeletal muscle (Micropogon opercularis). Int. J. Biochem. Cell Biol. 33: 691-699 https://doi.org/10.1016/S1357-2725(01)00054-1
  30. Sen, L. J. and J. R. Whiteker. 1973. Some properties of a ficin-papain inhibitor from avian egg white. Arch. Biochem. Biophys. 158: 623-632 https://doi.org/10.1016/0003-9861(73)90554-7
  31. Sharekar, S. V., M. S. Gore, and V. Ninjoor. 1988. Purification and characterization of cathepsin B from skeletal muscle of fresh water fish, Tilapia mossambica. J. Food Sci. 53: 1018-1023 https://doi.org/10.1111/j.1365-2621.1988.tb13521.x
  32. Sueyoshi, T., K. Enjyoji, T. Shimada, H. Kato, S. Iwanaga, Y. Bando, E. Kominami, and N. Katunuma. 1985. A new function of kininogens as thiol-proteinase inhibitor: Inhibition of papain and cathepsins B, H, and L by bovine, rat and human plasma kininogen. FEBS 182: 193-195 https://doi.org/10.1016/0014-5793(85)81182-0
  33. Synnes, M. 1998. Purification and characterization of two cysteine proteinase inhibitors from the skin of Atlantic salmon (Salmo salar L.). Comp. Biochem. Physiol. B 121: 257-264 https://doi.org/10.1016/S0305-0491(98)10098-6
  34. Takio, K., E. Kaminami, N. Wakamatsu, N. Katunuma, and K. Titani. 1983. Amino acid sequence of rat liver proteinases inhibitor. Biochem. Biophys. Res. Commun. 115: 902-908 https://doi.org/10.1016/S0006-291X(83)80020-5
  35. Toyohara, H., Y. Makinodan, and S. Ikeda. 1988. Detection of a cysteine protease inhibitor in carp muscle. Nippon Suisan Gakkaishi 54: 157
  36. Tsai, Y. J., G. D. Chang, C. J. Huang, Y. S. Chang, and F. L. Huang. 1996. Purification and molecular cloning of carp ovarian cystatin. Comp. Biochem. Physiol. B 113: 573- 580 https://doi.org/10.1016/0305-0491(95)02070-5
  37. Turk, B., I. Kriza, and V. Turk. 1992. Isolation and characterization of bovine stefin B. Biol. Chem. Hoppe Seyler 373: 441-446 https://doi.org/10.1515/bchm3.1992.373.2.441
  38. Turk, B., V. Stoka, V. Turk, G. Johansson, J. J. Cazzulo, and I. Bjork. 1996. High-molecular-weight kininogen binds two molecules of cysteine proteinases with different rate constants. FEBS Lett. 391: 109-112 https://doi.org/10.1016/0014-5793(96)00611-4
  39. Ustadi, K. Y. Kim, and S. M. Kim. 2005. Purification and identification of a protease inhibitor from Glassfish (Liparis tanakai) Eggs. J. Agric. Food Chem. 53: 7667-7672 https://doi.org/10.1021/jf0482459
  40. Walker, J. M. 2002. Nondenaturing polyacrylamide gel electrophoresis of protein, pp. 57-60. In J. M. Walker (ed.), The Protein Protocols Handbook, 2nd Ed. Huana Press, Totowa, NJ, U.S.A
  41. Weerasinghe V. C., M. T. Morrissey, and H. An. 1996. Characterization of active components in food grade proteinase inhibitor for surimi manufacture. J. Food Chem. 44: 2584- 2590 https://doi.org/10.1021/jf950589z
  42. Bae, E.-A., J.-E. Shin, S.-H. Park, and D.-H. Kim. 2004. Inhibitory effect of ginseng polysaccharides on rotavirus infection. J. Microbiol. Biotechnol. 14: 202-204
  43. Yamashita, M. and S. Konagaya. 1990. Purification and characterization of cathepsin L from the white muscle of chum salmon, Oncorhynchus keta. Comp. Biochem. Physiol. 96B: 247-252 https://doi.org/10.1016/0305-0491(90)90371-Y
  44. Yamashita, M. and S. Konagaya. 1990. Purification and characterization of cathepsin B from the white muscle of chum salmon, Oncorhynchus keta. Comp. Biochem. Physiol. 96B: 733-737 https://doi.org/10.1016/0305-0491(90)90222-F
  45. Yamashita, M. and S. Konagaya. 1991. Cysteine protease inhibitor in egg of chum salmon. J. Biochem. 110: 762-766 https://doi.org/10.1093/oxfordjournals.jbchem.a123655
  46. Yamashita, M. and S. Konagaya. 1991. A comparison of cystatin activity in various tissues of chum salmon (Oncorhynchus keta) between feeding and spawning migrations. Comp. Biochem. Physiol. A 100: 749-751 https://doi.org/10.1016/0300-9629(91)90402-X
  47. Ylonen, A., A. Rinne, J. Herttuainen, J. Bogwald, M. Jarvinen, and N. Kalkkinen. 1999. Atlantic salmon (Salmo salar L) skin contains a novel kininogen and another cysteine proteinase inhibitor. Eur. J. Biochem. 266: 1066-1072 https://doi.org/10.1046/j.1432-1327.1999.00950.x