Skip to main content
SpringerLink
Log in

Purification and characterization of lysozyme from filipino venus, Ruditapes philippinarum

  • Research Article
  • Published:
Food Science and Biotechnology Aims and scope Submit manuscript

Abstract

Lysozyme from Filipino venus (Ruditapes philippinarum) was purified by ion-exchange and gel filtration chromatography. The purification fold and yield were 3,402 and 32.4%, respectively. The molecular weight was determined to be 13.4 kDa by SDS-PAGE. The specific activity of lysozyme was 3.76×105 units/mg protein with Micrococcus lysodeikticus as a substrate. The optimum temperature and pH of lysozyme were 75°C and 5.5, respectively. Lysozyme activity was decreased with about 45% after heat treatment for 30 min at 80°C, and completely inactivated at 100°C. It was activated by NaCl (10–70 mM), MgCl2, and CaCl2 (2–5 mM) whereas it was inhibited by ZnCl2 (2–30 mM).

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Jolles P, Jolles E. What’s new in lysozyme research? Always a model system, today as yesterday. Mol. Cell. Biochem. 63: 165–189 (1984)

    Article  CAS  Google Scholar 

  2. Salton MRJ. The properties of lysozyme and its action on microorganisms. Bacteriol. Rev. 21: 82–99 (1957)

    CAS  Google Scholar 

  3. Ibrahim HR, Thomas U, Pellegrini A. A helix-loop-helix-peptide at the upper lip of the active site cleft of lysozyme confers potent antimicrobial activity with membrane permeabilization action. J. Biol. Chem. 276: 43767–43774 (2001)

    Article  CAS  Google Scholar 

  4. Kornegay JR, Schilling JW, Wilson AC. Molecular adaptation of a leaf-eating bird: Stomach lysozyme of the hoatzin. Mol. Biol. Evol. 11: 921–928 (1994)

    CAS  Google Scholar 

  5. Datta S, Janes ME, Xue Q-G, Losso J, La Peyre JF. Control of Listeria monocytogenes and Salmonella anatum on the surface of smoked salmon coated with calcium alginate coating containing oyster lysozyme and nisin. J. Food Sci. 73: M67–M71 (2008)

    Article  CAS  Google Scholar 

  6. Mecitoglu C, Yemenicioglu A, Arslanoglu A, Elmaci ZS, Korel F, Cetin AE. Incorporation of partially purified hen egg white lysozyme into zein films for antimicrobial food packaging. Food Res. Int. 39: 12–21 (2006)

    Article  CAS  Google Scholar 

  7. Min S, Harris LJ, Han JH, Krochta JM. Listeria monocytogenes inhibition by whey protein films and coatings incorporating lysozyme. J. Food Protect. 68: 2317–2325 (2005)

    CAS  Google Scholar 

  8. Ito Y, Yoshikawa A, Hotanii T, Fukuda S, Sugimura K, Imoto T. Amino acid sequences of lysozymes newly purified from invertebrates imply wide distribution of a novel class in the lysozyme family. Eur. J. Biochem. 259: 456–461 (1999)

    Article  CAS  Google Scholar 

  9. Xue QG, Schey KL, Volety AK, Chu FL, La Peyre JF. Purification and characterization of lysozyme from plasma of the eastern oyster (Crassostrea virginica). Comp. Biochem. Physiol. 139: 11–25 (2004)

    Google Scholar 

  10. McHenery JG, Birbeck TH. Characterization of the lysozyme of Mytilus edulis (L). Comp. Biochem. Physiol. B 71: 583–589 (1982)

    Article  CAS  Google Scholar 

  11. Myrenes B, Johansen A. Recovery of lysozyme from scallop waste. Prep. Biochem. 24: 69–80 (1994)

    Article  Google Scholar 

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

    Article  CAS  Google Scholar 

  13. Shugar D. Measurement of lysozyme activity and the ultra violet inactivation of lysozyme. Biochem. Biophys. Acta 8: 302–309 (1952)

    Article  CAS  Google Scholar 

  14. Laemmli UK. Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 227: 680–685 (1970)

    Article  CAS  Google Scholar 

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

    CAS  Google Scholar 

  16. Lee JM, Kim SM, Kim SM. Biochemical and antibacterial properties of lysozyme purified from the viscera of scallops (Patinopecten yessoensis). J. Food Biochem. 32: 474–489 (2008)

    Article  CAS  Google Scholar 

  17. Thammasirirak S, Ponkhan P, Preecharram S, Khanchanuan R, Phonyothee P, Daduang S, Srisomsap C, Araki T, Svasti J. Purification, characterization, and comparison of reptile lysozymes. Comp. Biochem. Physiol. 143: 209–217 (2006)

    Article  Google Scholar 

  18. Lockey TD, Ourth DD. Purification and characterization of lysozyme from hemolymph of Heliothis virescens larvae. Biochem. Bioph. Res. Co. 220: 502–508 (1996)

    Article  CAS  Google Scholar 

  19. Schoentgen F, Jolles J, Jolles P. Complete amino acid sequence of ostrich egg-white lysozyme, a goose-type lysozyme. Eur. J. Biochem. 123: 489–497 (1982)

    Article  CAS  Google Scholar 

  20. Nilsen IW, Overbo K, Sandsdalen E, Sandaker E, Sletten K, Bjornar M. Protein purification and gene isolation of chlamysin, a cold active lysozyme-like enzyme with antibacterial activity. FEBS Lett. 464: 153–158 (1999)

    Article  CAS  Google Scholar 

  21. Miyauchi K, Matsumiya M, Mochizuki A. Purification and characterization of lysozyme from purple Washington clam Saxidomus purpurata. Fisheries Sci. 72: 1300–1305 (2000)

    Article  Google Scholar 

  22. Wang S, Ng TB, Chen T, Lin D, Wu J, Rao P, YE X. First report of a novel plant lysozyme with both antifungal and antibacterial activities. Biochem. Biophys. 327: 820–827 (2005)

    Article  CAS  Google Scholar 

  23. Miyauchi K, Matsumiya M, Mochizuki A. Purification and characterization of lysozyme from brackishwater clam Corbicula japonica. Nippon Suisan Gakk. 66: 275–281 (2000)

    Article  CAS  Google Scholar 

  24. Wang S, Ye X, Rao P. Isolation of a novel leguminous lysozyme and study on the antifungal activity. Food Res. Int. 47: 341–347 (2012)

    Article  CAS  Google Scholar 

  25. Minagawa S, Hikima J, Hirono I, Aoki T. Expression of Japanese flounder cDNA in insect cells. Dev. Comp. Immunol. 25: 439–445 (2001)

    Article  CAS  Google Scholar 

  26. Grinde B, Jolles J, Jolles P. Purification and characterization of two lysozymes from rainbow trout (Salmo gairdneri). Eur. J. Biochem. 173: 269–273 (1988)

    Article  CAS  Google Scholar 

  27. Fujimoto S, Toshimori-Tsuda I, Kishimoto K, Yamano Y, Morishima I. Protein purification, cDNA cloning, and gene expression of lysozyme from eri-silkworm, Samia Cynthia ricini. Comp. Biochem. Phys. B 128: 709–718 (2001)

    Article  CAS  Google Scholar 

  28. Hikima S, Hikima J, Rojtinnakorn J, Hirono I, Aoki T. Characterization and function of kuruma shrimp lysozyme possessing lytic activity against Vibrio species. Gene 316: 187–195 (2003)

    Article  CAS  Google Scholar 

  29. Klomklao S, Benjakul S, Visessanguan W, Kishimura H, Simpson BK, Saeki H. Trypsins from yellowfin tuna (Thunnus albacores) spleen: Purification and characterization. Comp. Biochem. Physiol. B 144: 47–56 (2006)

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Food Science and Nutrition, Dankook University, Yongin, Gyeonggi, 448-701, Korea

    Misook Kim, Minjeong Park & Yoonhwa Jeong

Authors
  1. Misook Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Minjeong Park
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yoonhwa Jeong
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yoonhwa Jeong.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kim, M., Park, M. & Jeong, Y. Purification and characterization of lysozyme from filipino venus, Ruditapes philippinarum . Food Sci Biotechnol 21, 1463–1468 (2012). https://doi.org/10.1007/s10068-012-0193-z

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10068-012-0193-z

Keywords

Search

Navigation