Abstract
Lysozyme from equine neutrophil granulocytes was isolated in a pure form by fast performance liquid chromatography, i.e. ion-exchange chromatography and reversed-phase chromatography. The lysozyme lysed Micrococcus luteus, Bacillus subtilis and Staphylococcus lentus and was also bactericidal against the Gram-negative bacteria Escherichia coli, Klebsiella pneumoniae, Bordetella bronchiseptica, and Serratia marcescens. Staphylococcus aureus and Staphylococcus epidermidis were not lysed. The lysozyme was only very slightly bactericidal for S. epidermidis and S. aureus. Equine neutrophil lysozyme was found to be bactericidal for Gram-positive as well as for Gram-negative bacteria without further treatment. Equine and chicken egg white lysozymes were found to be immunologically related when examined using specific antisera against each of them. Both lysozymes also had very similar specific enzymatic activities against M. luteus membranes.
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Eisenhauer, P.B., Harwig, S.S.L., Szklarek, D., Ganz, T., Selsted, M.E. and Lehrer R.I., 1989. Purification and antimicrobial properties of three defensins from rat neutrophils. Infection and Immunity, 57, 2021–2027
Feingold, D.S., Goldmann, J. and Kuritz, H.M., 1968. Locus of the action of serum and the role of lysozyme in the serum bactericidal reaction. Journal of Bacteriology, 96, 2118–2126
Ganz, T., Selsted, M.E., Szklarek, D., Harwig, S.S.L., Daher, K., Bainton, D.F. and Lehrer, R.I., 1985. Defensins: Natural peptide antibiotics of human neutrophils. Journal of Clinical Investigation, 76, 1427–1435
Jollès, P. and Jollès, J., 1984. What's new in lysozyme research? Molecular and Cellular Biochemistry, 63, 165–189
Laemmli, U.K., 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage, T4. Nature, 227, 680–685
Lehrer, R.I., Szklarek, D., Ganz, T. and Selsted, M.E., 1986. Synergic activity of rabbit granulocyte peptides against Candida albicans. Infection and Immunity, 52, 902–904
Ouchterlony, O. and Nilson, L.A., 1978. In: D.M.Weir (ed.), Handbook of Experimental Immunology, (Blackwells, London) 19.1–19.44
Padgett, G.A. and Hirsch, J.G., 1967. Lysozyme: its absence in tears and leukocytes of cattle. Australian Journal of Experimental Biology and Medical Science, 45, 569–570
Pellegrini, A., Hägeli, G. and vonFellenberg, R., 1988. Isolation and characterization of two new low molecular weight protein proteinase inhibitors from the granular-rich fraction of equine neutrophilic granulocytes. Biochimica et Biophysica Acta, 952, 309–316
Pellegrini, A., Grob, K. and vonFellenberg, R., 1990. Identification of a bactericidal component active against Gram-positive and Gram-negative bacteria in commercial ovomucoid and ovoinhibitor as lysozyme. Letters in Applied Microbiology, 10, 201–204
Pellegrini, A., Thomas, U and von Fellenberg, R., 1991. Bactericidal activities of lysozyme and aprotinin against Gram-negative and Gram-positive bacteria related to their basic character. Journal of Applied Bacteriology, in press
Rausch, P. and Moore, T.G., 1975. Granule enzymes of polymorphonuclear neutrophils: A phylogenetic comparison. Blood, 46, 913–919
Repaske, R., 1958. Lysis of Gram-negative organisms and the role of versene. Biochimica et Biophysica Acta, 30, 225–232
Spitznagel, J.K., 1984. Non-oxidative antimicrobial reactions of leukocytes. Contemporary Topics in Immunology, 14, 283–343
Thorne, K.J.J., Oliver, R.C. and Barrett, A.J., 1976. Lysis and killing of bacteria by lysosomal proteinases. Infection and Immunity, 14, 555–563
Waiblinger, S., 1990. Reinigung und partielle Charakterisierung von Lysozym und antibakteriellen Proteinen aus neutrophilen Granulozyten vom Pferd, (Doctoral dissertation, Zurich)
Wilson, L.A. and Spitznagel, J.K., 1968. Molecular and structural damage of Escherichia coli produced by antibody, complement and lysozyme system. Journal of Bacteriology, 96, 1339–1348
Yamashita, T. and Saito, K., 1989. Purification, primary structure and biological activity of guinea pig neutrophil cationic peptides. Infection and Immunity, 57, 2405–2409
Zerial, A., Skezlavaj, B., Gennaro, R. and Romeo, D., 1987. Inactivation of herpes simplex virus by protein component of bovine neutrophil granules. Antiviral Research, 7, 341–352
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Pellegrini, A., Waiblinger, S. & von Fellenberg, R. Purification of equine neutrophil lysozyme and its antibacterial activity against Gram-positive and Gram-negative bacteria. Veterinary Research Communications 15, 427–435 (1991). https://doi.org/10.1007/BF00346538
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DOI: https://doi.org/10.1007/BF00346538