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Applied and Environmental Microbiology, February 1999, p. 569-577, Vol. 65, No. 2
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Identification and Characterization of a Lysis Module Present in a Large Proportion of Bacteriophages Infecting Streptococcus thermophilus

Michelle M. Sheehan,1 Elizabeth Stanley,1 Gerald F. Fitzgerald,1,2,3 and Douwe van Sinderen2,*

The National Food Biotechnology Centre1 and Departments of Microbiology2 and Food Science and Technology,3 University College Cork, Cork, Ireland

Received 27 April 1998/Accepted 29 October 1998

A lysis module encoded by the temperate bacteriophage phi O1205 was identified. This lysis module contains a lysin gene, designated lyt51, and two putative holin-encoding genes, designated lyt49 and lyt50. lyt51 encodes a lytic enzyme specifically directed against streptococcal cell walls. Similar to other phage-encoded lysins, Lyt51 appears to have a modular design in which the N-terminal portion corresponds to its enzymatic activity while the C-terminal region is responsible for its substrate binding specificity. The two putative holin-encoding genes, lyt49 and lyt50, located immediately upstream of lyt51, were identified on the basis of their homology to other identified holin-encoding genes. Expression of lyt49 or lyt50 in Escherichia coli was shown to cause cell death and leakage of the intracellular enzyme isocitrate dehydrogenase into the growth medium without apparent lysis of the cells. Southern blotting experiments demonstrated that at least one of the three components of the identified lysis module is present in all members of a large collection of bacteriophages, indicating that components of this lysis module are widespread among bacteriophages infecting Streptococcus thermophilus.

* Corresponding author. Mailing address: Department of Microbiology, University College Cork, Cork, Ireland. Phone: 353 21 902811. Fax: 353 21 903101. E-mail: douwe{at}ucc.ie.

Applied and Environmental Microbiology, February 1999, p. 569-577, Vol. 65, No. 2
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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