Volume 8, Issue 2 (2020)                   ECOPERSIA 2020, 8(2): 125-131 | Back to browse issues page

XML Print


Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:

Ghalambor M, Eslamifar Z, Khoshnood Z. Biochemical Characterization of Lysozyme Extracted from Common Carp, Cyprinus carpio. ECOPERSIA 2020; 8 (2) :125-131
URL: http://ecopersia.modares.ac.ir/article-24-36520-en.html
1- Biology Department, Science Faculty, Dezful Branch, Islamic Azad University, Dezful, Iran
2- Biochemistry Department, Medical Faculty, Dezful University of Medical Sciences, Dezful, Iran
3- Biology Department, Science Faculty, Dezful Branch, Islamic Azad University, Dezful, Iran , zkhoshnood@gmail.com
Abstract:   (2449 Views)
Aim: The aim of the present study was to investigate the biochemical characterizations of the lysozyme enzyme for evaluation of its importance in the immune system of the common carp, Cyprinus carpio.
Materials & Methods: In the present study, lysozyme was extracted from the spleen of common carp, Cyprinus carpio. Then, partially purified by ammonium sulfate and some properties such as optimum pH and temperature as well as the effects of different salt concentrations of NaCl, MgCl2, KCl, and urea on enzyme activity were evaluated. The enzyme activity was assayed using a suspension of Micrococcus lysodeikticus as a substrate.
Findings: The optimum pH and temperature were found 4 and 50°C, respectively. Furthermore, lysozyme activity was found to be dependent on salt concentration.
Conclusion: Based on the results, it's been concluded that lysozyme extracted from the spleen of the C. carpio has its optimum activity at high temperature and low pH condition and its activity could be continued with the presence of different salt compounds which all these are related to the environmental conditions of natural habitats of the C. carpio and showed that lysozyme could be one of the key factors of the immune system in this species.
 
Full-Text [PDF 431 kb]   (1218 Downloads)    
Article Type: Original Research | Subject: Aquatic Ecology
Received: 2019/09/16 | Accepted: 2019/12/26 | Published: 2020/05/19
* Corresponding Author Address: Biology Department, Dezful Branch, Islamic Azad University, Azadegan, University Boulevard, Dezful, Khuzestan, Iran

References
1. Uribe C, Folch H, Enriquez R, Moran G. Innate and adaptive immunity in teleost fish: A review. Vet Med. 2011;56(10):486-503. [Link] [DOI:10.17221/3294-VETMED]
2. Bowden TJ. Modulation of the immune system of fish by their environment. Fish Shellfish Immunol. 2008;25(4):373-83. [Link] [DOI:10.1016/j.fsi.2008.03.017]
3. Gao Ch, Fu Q, Zhou Sh, Song L, Ren Y, Dong X, et al. The mucosal expression signatures of g-type lysozyme in turbot (Scopthalmus maximus) following bacteria challenge. Fish Shellfish Immunol. 2016;54:612-19. [Link] [DOI:10.1016/j.fsi.2016.05.015]
4. Subramanian S, Ross NW, MacKinnon SL. Comparison of antimicrobial activity in the epidermal mucus extracts of fish. Comp Biochem Physiol Part B Biochem Mol Biol. 2008;150(1):85-92. [Link] [DOI:10.1016/j.cbpb.2008.01.011]
5. Fletcher T. Defense mechanisms in fish. In: Malins DC, Sargent JR. Biochemical and biophysical perspectives in marine biology. Cambridge: Academic Press; 1978. pp. 122-89. [Link]
6. Ibrahim H, Yamada M, Kobayashi K. Bactericidal action of lysozyme against gram-negative bacteria due to insertion of a hydrophobic pentapeptide into its C-terminus. Biosci Biotechnol Biochem. 1992;56(8):1361-3. [Link] [DOI:10.1271/bbb.56.1361]
7. Pickering A. The distribution of mucus cells in the epidermis of the brown trout Salmo trutta (L.) and the char Salvelinus alpinus (L). J Fish Biol. 1974;6(2):111-8. [Link] [DOI:10.1111/j.1095-8649.1974.tb04531.x]
8. Shepherd KL. Functions for fish mucus. Rev Fish Biol Fish. 1994;4(4):401-29. [Link] [DOI:10.1007/BF00042888]
9. Cole AM, Weis P, Diamond G. Isolation and characterization of pleurocidin, an antimicrobial peptide in the skin secretions of winter flounder. J Biol Chem. 1997;272(18):12008-13. [Link] [DOI:10.1074/jbc.272.18.12008]
10. Gomez GD, Balcazar JL. A review on the interactions between gut microbiota and innate immunity of fish. FEMS Immunol Med Microbiol. 2008;52(2):145-54. [Link] [DOI:10.1111/j.1574-695X.2007.00343.x]
11. Grove S, Johansen R, Reitan L, Press CM. Immune- and enzyme histochemical characterisation of leukocyte populations within lymphoid and mucosal tissues of Atlantic halibut (Hippoglossus hippoglossus). Fish Shellfish Immunol. 2006;20(5):693-708. [Link] [DOI:10.1016/j.fsi.2005.08.009]
12. Biller-Takahashi JD, Urbinati EC. Fish Immunology. The modification and manipulation of the innate immune system: Brazilian studies. Anais da Academia Brasileira de Ciências. 2014;86(3):1484-506. [Link] [DOI:10.1590/0001-3765201420130159]
13. Magnadottir B. Innate immunity of fish (overview). Fish Shellfish Immunol. 2006;20(2):137-51. [Link] [DOI:10.1016/j.fsi.2004.09.006]
14. Zapata A, Diez B, Cejalvo T, Gutiérrez-de Frías C, Cortés A. Ontogeny of the immune system of fish. Fish Shellfish Immunol. 2006;20(2):126-36. [Link] [DOI:10.1016/j.fsi.2004.09.005]
15. Zapata AG, Torroba M, Varas A, Jimenez AV. Immunity in fish larvae. Dev Biol Stand. 1997;90:23-32. [Link]
16. Jolles P, Jolles E. What's new in lysozyme research?. Mol Cell Biochem. 1984;63(2):165-89. [Link] [DOI:10.1007/BF00285225]
17. Masschalck B, Michiels CW. Antimicrobial properties of lysozyme in relation to foodborne vegetative bacteria. Crit Rev Microbiol. 2003;29(3):191-214. [Link] [DOI:10.1080/713610448]
18. Zhao J, Song L, Li Ch, Zou H, Ni D, Wang W, et al. Molecular cloning of an invertebrate goose-type lysozyme gene from Chlamys farreri, and lytic activity of the recombinant protein. Mol Immunol. 2007;44(6):1198-208. [Link] [DOI:10.1016/j.molimm.2006.06.008]
19. Sotelo-Mundo RR, Islas-Osun MA, De-Lare-Vega E, Hernandez-Lopez J, Valgasalbores F, Yepiz-Plascencia G. cDNA cloning of the lysozyme of the white shrimp Penaeus vannamei. Fish Shellfish Immunol. 2003;15(4):325-31. [Link] [DOI:10.1016/S1050-4648(02)00176-6]
20. Guani-Guerra E, Santos-Mendoza T, Lugo-Reyes SO, Teran LM. Antimicrobial peptides: General overview and clinical implications in human health and disease. Clin Immunol. 2010;135(1):1-11. [Link] [DOI:10.1016/j.clim.2009.12.004]
21. Tassanakajon A, Somboonwiwat K, Supungul P, Tang S. Discovery of immune molecules and their crucial functions in shrimp immunity. Fish Shellfish Immunol. 2013;34(4):954-67. [Link] [DOI:10.1016/j.fsi.2012.09.021]
22. Modanloo M, Soltanian S, Akhlaghi M, Hoseinifar SH. The effects of single or combined administration of galactooligosaccharide and Pediococcus acidilactici on cutaneous mucus immune parameters, humoral immune responses and immune related genes expression in common carp (Cyprinus carpio) fingerlings. Fish Shellfish Immunol. 2017;70:391-7. [Link] [DOI:10.1016/j.fsi.2017.09.032]
23. Yano T. The non-specific immune system: Humoral defense. In: Iwama G, Nakanishi T, editors. The fish immune system: Organism, pathogen, and environment. Cambridge: Academic Press; 1996. pp. 106-57. [Link] [DOI:10.1016/S1546-5098(08)60273-3]
24. Soltani M, Sheikhzadeh N, Ebrahimzadeh-Mousavi HA, Zargar A. Effect of Zatana mutiflora essential oil on innate immune responses of common carp (Cyprinus carpio). J Fish Aquat Sci. 2010;5(3):191-9. [Link] [DOI:10.3923/jfas.2010.191.199]
25. Bazrkar V, Aghamaali MR. Biochemical characterization of lysozyme from of Rutilus frisii kutum. Aquat Physiol Biotechnol. 2015;2(4):23-34. [Persian] [Link]
26. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;227(5259):680-5. [Link] [DOI:10.1038/227680a0]
27. Minagawa S, Hikima J, Hirono I, Aoki T. Expression of Japanese flounder c-type lysozyme cDNA in insect cells. Dev Comp Immunol. 2001;25(5-6):439-45. [Link] [DOI:10.1016/S0145-305X(01)00013-1]
28. Grinde B, Jolles J, Jolles P. Purification and characterization of two lysozymes from rainbow trout (Salmo gairdneri). Eur J Biochem. 1988;173(2):269-73. [Link] [DOI:10.1111/j.1432-1033.1988.tb13994.x]
29. 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. 1999;259(1-2):456-61. [Link] [DOI:10.1046/j.1432-1327.1999.00064.x]
30. Nilsen IW, Overbo K, Sandsdalen E, Sandaker E, Sletten K, Myrnes B. Protein purification and gene isolation of chlamysin, a cold-active lysozyme-like enzyme with antibacterial activity. FEBS Lett. 1999;464(3):153-8. [Link] [DOI:10.1016/S0014-5793(99)01693-2]
31. Xue QG, Schey KL, Volety AK, Chu FL, La Peyere JF. Purification and characterization of lysozyme from plasma of the eastern oyster (Crassostrea virginea). Comp Biochem Physiol Part B Biochem Mol Biol. 2004;139(1):11-25. [Link] [DOI:10.1016/j.cbpc.2004.05.011]
32. McHenery JG, Birbeck TH. Characterization of the lysozyme of Mytilus edulis (L). Comp Biochem Physiol B Comp Biochem. 1982;71(4):583-9. [Link] [DOI:10.1016/0305-0491(82)90466-7]
33. Wang Sh, Ye X, Rao P. Isolation of a novel leguminous lysozyme and study on the antifungal activity. Food Res Int. 2012;47(2):341-7. [Link] [DOI:10.1016/j.foodres.2011.07.039]
34. Wang Sh, Ng TB, Chen T, Lin D, Wu J, Rao P, et al. First report of a novel plant lysozyme with both antifungal and antibacterial activities. Biochem Biophys Res Commun. 2005;327(3):820-7. [Link] [DOI:10.1016/j.bbrc.2004.12.077]
35. Lee JM, Kim SM, Kim SM. Biochemical and antibacterial properties of lysozyme purified from the viscera of scallops (Patinopecten yessoensis). J Food Biochem. 2008;32(4):474-89. [Link] [DOI:10.1111/j.1745-4514.2008.00177.x]
36. 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 Physiol Part B Biochem Mol Biol. 2001;128(4):709-18. [Link] [DOI:10.1016/S1096-4959(00)00368-7]
37. Hikima S, Hikima JI, Rojtinnakorn J, Hirono I, Aoki T. Characterization and function of kuruma shrimp lysozyme possessing lytic activity against Vibrio species. Gene. 2003;316:187-95. [Link] [DOI:10.1016/S0378-1119(03)00761-3]
38. Kim M, Park M, Jeong Y. Purification and characterization of lysozyme from fillipino venus, Ruditapes philippinarum. Food Sci Biotechnol. 2012;21(5):1463-8. [Link] [DOI:10.1007/s10068-012-0193-z]
39. 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 Part-B Biochem Mol Biol. 2006;144(1):47-56. [Link] [DOI:10.1016/j.cbpb.2006.01.006]
40. Sidhan V, Gurnani S. Kinetic characterization of rat liver nuclear lysozyme. J Biosci. 1982;4(2):191-5. [Link] [DOI:10.1007/BF02702729]
41. Emadi S, Behzadi M. A comparative study on the aggregating effects of guanidine thiocyanate, guanidine hydrochloride and urea on lysozyme aggregation. Biochem Biophys Res Commun. 2014;450(4):1339-44. [Link] [DOI:10.1016/j.bbrc.2014.06.133]

Add your comments about this article : Your username or Email:
CAPTCHA

Send email to the article author


Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.