Streptococcus thermophilus autolytic strains are characterized by a typical bell-shaped growth curve when grown under appropriate conditions. The cellular mechanisms involved in the triggering of lysis and the bacteriolytic activities of these strains were investigated in this study. Lactose depletion and organic solvents (ethanol, methanol, and chloroform) were shown to trigger a premature and immediate lysis of M17 exponentially growing cells. These factors and compounds are suspected to act by altering the cell envelope properties, causing either the permeabilization (organic solvents) or the depolarization (lactose depletion) of the cytoplasmic membrane. The autolytic character was shown to be associated with lysogeny. Phage particles, most of which were defective, were observed in the culture supernatants after both mitomycin C-induced and spontaneous lysis. By renaturing sodium dodecyl sulfate-polyacrylamide gel electrophoresis, a bacteriolytic activity was detected at 31 kDa exclusively in the autolytic strains. This enzyme was detected during both growth and spontaneous lysis with the same intensity. We have shown that it was prophage encoded and homologous to the endolysin Lyt51 of the streptococcal temperate bacteriophage 01205 (M. Sheehan, E. Stanley, G. F. Fitzgerald, and D. van Sinderen, Appl. Environ. Microbiol. 65:569-577, 1999). It appears from our results that the autolytic properties are conferred to the S. thermophilus strains by a leaky prophage but do not result from massive prophage induction. More specifically, we propose that phagic genes are constitutively expressed in almost all the cells at a low and nonlethal level and that lysis is controlled and achieved by the prophage-encoded lysis proteins.Autolysis of lactic acid bacteria used as starters appears to be a crucial step in the flavor development of fermented dairy products (12,15). It indeed causes the opening of cells and the subsequent release into the curd of intracellular enzymes that are involved in the flavor compound formation. The activity of these enzymes can therefore be enhanced by better accessibility to their substrates, proteins, lipids, or carbohydrates from the milk. It has been shown that the main consequence of cell autolysis in cheese is peptidolysis acceleration, leading to an increased rate of free amino acid production and to a decrease in bitter taste (2,3,13,16,25,32,34,35,55).Bacterial autolysis results from the degradation of the peptidoglycan, which is the major structural component of the bacterial cell wall, by enzymes called peptidoglycan hydrolases. Bacteria synthesize their own peptidoglycan hydrolases, named autolysins (45). These cell wall-associated enzymes are potentially lethal for the cell and thus require stringent regulation. It has been proposed that autolysins are involved in different cellular processes including cell wall expansion and turnover, cell division, and transformation (47). Autolysis would result from an uncontrolled action of the bacterial autolysins after inhibition of peptid...