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Partial purification and some properties of øC2(W) lysin, a lytic enzyme produced by phage-infected cells of Streptococcus lactis C2

Published online by Cambridge University Press:  01 June 2009

W. Michael A. Mullan  and
Robert J. M. Crawford
W. Michael A. Mullan
Affiliation:
Department of Dairy Technology, West of Scotland Agricultural College, AyrScotland
Robert J. M. Crawford
Affiliation:
Department of Dairy Technology, West of Scotland Agricultural College, AyrScotland
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Summary

The lytic enzyme present in øC2(W) lysates was isolated by means of ion-exchange chromatography and further purified by gel filtration and ultrafiltration. The phage enzyme had an apparent pH optimum of 6·5–6·9 and was rapidly inactivated at temperatures > 47°C. The apparent temperature optimum was 37°C and Q10 and Ea values over the range 22–32°C were 2·5 and 69·2 kJ/mol respectively. Monovalent and divalent cations activated the enzyme. Reduced -SH groups on the enzyme were required for lytic activity. Gel filtration revealed a mol. wt of ˜ 46000. Strain-dependent differences in sensitivity of group N lactic streptococci to lysin were found. Group D streptococci were also lysed. Strains of three species of Leuconostoc, two species of Lactobacillus, one strain of Escherichia coli and of Micrococcus lysodeikticus were apparently resistant. Analysis of cell wall degradation products gave results which were consistent with the lysin having the specificity of an N-acetylmuramidase.

Type
Original Articles
Information
Journal of Dairy Research , Volume 52 , Issue 1 , February 1985 , pp. 123 - 138
Copyright
Copyright © Proprietors of Journal of Dairy Research 1985

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References

REFERENCES

Andrews, P. 1964 Estimation of the molecular weights of proteins by Sephadex gel-filtration. Biochemical Journal 91 222–233CrossRefGoogle ScholarPubMed
Barkulis, S. S., Smith, C., Botralik, J. J. & Heyman, H. 1964 Structure of streptococcal cell walls. IV. Purification and properties of streptococcal phage muralysin. Journal of Biological Chemistry 239 4027–4033CrossRefGoogle ScholarPubMed
Bradford, M. M. 1976 A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72 248–254CrossRefGoogle ScholarPubMed
Brumfitt, W., Wardlaw, A. C. & Park, J. T. 1958 Development of lysozyme-resistance in Micrococcus lysodeikticus and its association with an increased O-acetyl content of the cell wall. Nature 181 1783–1784CrossRefGoogle Scholar
Cleveland, R. F., Wioken, A. J., Daneo-Moore, L. & Shockman, G. D. 1976 Inhibition of wall autolysis in Streptococcus faecalis by lipoteichoic acids and lipids. Journal of Bacteriology 126 192–197CrossRefGoogle ScholarPubMed
Collins, E. B. 1952 Action of bacteriophage on mixed strain starter cultures. I. Nature and characteristics of the ‘nascent phenomenon’. Journal of Dairy Science 35 371–380CrossRefGoogle Scholar
De Man, J. C., Rooosa, M. & Sharpe, M. E. 1960 A medium for the cultivation of lactobacilli. Journal of Applied Bacteriology 23 103–105CrossRefGoogle Scholar
Dische, Z. 1962 Color reactions based on the reducing properties of sugars. Methods in Carbohydrate Chemistry 1 512–514Google Scholar
Dixon, M. & Webb, E. C. 1964 Enzymes, 2nd edn. London: Longmans, Green & Co LtdGoogle Scholar
Ghuysen, J. M., Tipper, D. J. & Strominoer, J. L. 1966 Enzymes that degrade bacterial cell walls. Methods in Enzymology 8 685–699CrossRefGoogle Scholar
Krause, R. M. & Mocarty, M. 1961 Studies on the chemical structure of the streptococcal cell wall. I. The identification of a mucopeptide in the cell walls of groups A and A variant streptococci. Journal of Experimental Medicine 114 127–140CrossRefGoogle Scholar
Mandelstam, M. H. & Strominoer, J. L. 1961 On the structure of the cell wall of Staphylococcus aureus (Copenhagen). Biochemical and Biophysical Research Communications 5 466–471CrossRefGoogle Scholar
Mou, L., Sullivan, J. J. & Jago, G. R. 1976 Autolysis of Streptococcus cremoris. Journal of Dairy Research 43 275–282CrossRefGoogle Scholar
Mullan, W. M. A. 1982 A study of bacteriophage with particular reference to a lysin-producing lactic streptococcal phage-host system. PhD Thesis. University of GlasgowGoogle Scholar
Mullan, W. M. A. & Crawford, R. J. M. 1982 Phage C2(W) lysin, a bacteriolytic enzyme produced by phage infected cells of Str. lactis C2. 21st International Dairy Congress, Moscow, 1 (2) 350–351Google Scholar
Mullan, W. M. A. & Crawford, R. J. M. 1985 Lysin production by øC2(W), a prolate phage for Streptococcus lactis C2. Journal of Dairy Research 52 113–121CrossRefGoogle Scholar
Mullan, W. M. A., Daly, C. & Fox, P. F. 1981 Effect of cheese-making temperatures on the interactions of lactic streptococci and their phages. Journal of Dairy Research 48 465–471CrossRefGoogle Scholar
Murphy, J. S. 1961 An agent derived from B. megaterium phage G which dissolves the bacterial cell wall. Virology 11 510–513CrossRefGoogle Scholar
Naylor, J. & Czulak, J. 1956 Host-phage relationship of cheese starter organisms. II. Effect of phage activity on heterologous strains of lactic streptococci. Journal of Dairy Research 23 126–130CrossRefGoogle Scholar
Oram, J. D. 1965 Studies on the effects of lactoperoxidase and of bacteriophage on lactic acid streptococci. PhD Thesis, University of ReadingGoogle Scholar
Oram, J. D. & Reiter, B. 1965 Phage-associated lysins affecting group N and group D streptococci. Journal of General Microbiology 40 57–70CrossRefGoogle Scholar
Pette, J. W. 1953 The action of bacteriophages on lactic acid bacteria. 13th International Dairy Congress The Hague 3 1180–1184Google Scholar
Ralston, D. J., Baer, B. S., Lieberman, M. & Krueoer, A. P. 1957 Lysis from without of S. aureus K, by the combined action of phage and virolysin. Journal of General Physiology 41 343–358CrossRefGoogle Scholar
Rogosa, M. 1970 Characters used in the classification of lactobacilli. International Journal of Systematic Bacteriology 20 519–533CrossRefGoogle Scholar
Schleifer, K. H. & Kandler, O. 1972 Peptidoglycan types of bacterial cell walls and their taxonomic implications. Bacteriological Reviews 36 407–477CrossRefGoogle ScholarPubMed
Shockman, G. D., Thompson, J. S. & Conover, M. J. 1967 The autolytic enzyme system of Streptococcus faecalis. IT. Partial characterization of the autolysin and its substrate. Biochemistry 6 1054–1065CrossRefGoogle ScholarPubMed
Spackman, D. H., Stein, W. H. & Moore, S. 1958 Automatic recording apparatus for use in the chromatography of amino acids. Analytical Chemistry 30 1190–1206CrossRefGoogle Scholar
Terzaghi, B. E. & Sandine, W. E. 1975 Improved medium for lactic streptococci and their bacteriophages. Applied Microbiology 29 807–813CrossRefGoogle ScholarPubMed
Tourville, D. R. & Tokuda, S. 1967 Lactic streptococcal phage-associated lysin. II. Purification and characterization. Journal of Dairy Science 50 1019–1024CrossRefGoogle Scholar
Tsuqita, A. 1971 Phage lysozyme and other lytic enzymes. In The Enzymes 3rd edn. Vol. 5, pp. 343–411. (Ed. Boyer, P. D.), New York: Academic Press IncGoogle Scholar
Wallinder, I. -B. & Neujahr, H. Y. 1971 Cell wall and peptidoglycan from Lactobacillus fermenti. Journal of Bacteriology 105 918–926CrossRefGoogle ScholarPubMed
Weaver, G. L., Krooer, M. & Katz, F. 1977 Deaminated chitin affinity chromatography: a method for the isolation, purification and concentration of lysozyme. Journal of Food Science 42 1084–1087CrossRefGoogle Scholar
Whitehead, H. R., Hunter, G. J. E. & Cox, G. A. 1952 Inhibition of bacterial growth by bacteriophage as distinct from lytic action. Journal of General Microbiology 6 21–29CrossRefGoogle ScholarPubMed