Mur-LH, the Broad-Spectrum Endolysin of Lactobacillus helveticus Temperate Bacteriophage φ-0303
| Autor: | Michel Piot, Stéphane Guezenec, Stéphanie-Marie Deutsch, Sylvie Lortal, Simon J. Foster |
|---|---|
| Rok vydání: | 2004 |
| Předmět: |
Lactobacillus casei
Molecular Sequence Data Lysin Genetics and Molecular Biology Biology Lactobacillus gasseri Applied Microbiology and Biotechnology Substrate Specificity Microbiology Bacteriophage Bacteriolysis Lysogenic cycle Endopeptidases Bacteriophages Amino Acid Sequence Cloning Molecular Lysogeny Lactobacillus helveticus Base Sequence Ecology food and beverages biology.organism_classification Recombinant Proteins Temperateness Lactobacillus Biochemistry Holin Food Science Biotechnology |
| Zdroj: | Applied and Environmental Microbiology. 70:96-103 |
| ISSN: | 1098-5336 0099-2240 |
| DOI: | 10.1128/aem.70.1.96-103.2004 |
| Popis: | The phenomenon of lysis in fermentation industries, especially the dairy industry, is very significant. Two mechanisms can be involved: either an autolysin, which is a bacterial enzyme capable of degrading cell wall peptidoglycan under specific conditions, or a bacteriophage (virulent or temperate). In the latter case, the last step of lysis involves two proteins: a holin, which is a transmembrane protein forming pores in the cytoplasmic membrane, and a lysin (endolysin), which hydrolyzes the peptidoglycan of the cell wall, reaching its substrate as a result of the pores formed. Some genes encoding phage endolysins have been characterized in lactic acid bacteria (LAB) (for a pre-1996 review, see reference 26). The available data concern the endolysins of lactococcal bacteriophages φ-vML3 (31, 32), φ-c2 (40), φ-LC3 (2), φ-Tuc2009 (1), POO1 (14), and φ10MC, a temperate bacteriophage of Oenococcus oeni (11). Regarding the bacteriophages of lactobacilli, the endolysins of phage LL-H of Lactobacillus delbrueckii (38), phages φ-gle (16) and SC921 (41) of Lactobacillus plantarum, phage mv1 of Lactobacillus bulgaricus (3), phage PL-1 of Lactobacillus casei (17), and phage φadh of Lactobacillus gasseri (13) have been studied. The biochemical activity has been clearly demonstrated for only a few of these enzymes (17, 38); sequence homologies suggest activity for the others. The endolysins of LAB have been shown to be organized into two domains: the N-terminal domain, which is responsible for the catalytic activity of the protein, and the C-terminal domain, which is responsible for the binding to the substrate. Lactobacillus helveticus is a species commonly used as starter for the production of Swiss-type cheeses. Up to now, no molecular work has been done to elucidate the working of the endolysin-encoding bacteriophage genes. The φ-0303 phage is a temperate phage of L. helveticus CNRZ 303 that can be induced after mitomycin C treatment of the strain or during the manufacture of Swiss-type cheese (9). It has an isometric head and a tail with a contractile sheath and belongs to Bradley group A (4) or to the Myoviridae family of the International Committee on Taxonomy of Viruses (24). It also belongs to the most important group of phages described in L. helveticus: the short-tailed group (30). Its DNA is 39.9 kb long (10). In this work, we describe the cloning of the φ-0303 phage lysin gene and its nucleotide sequence, as well as the spectrum of activity and specificity of the recombinant protein. |
| Databáze: | OpenAIRE |
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