Mechanism of Nitrite-Induced Germination of Clostridium perfringens Spores

A study has been undertaken to understand the mechanism(s) of the nitrite-induced germination of Clostridium perfringens S40 spores. An increase in germination rates of the spores in response to increasing NaNO2 concentrations was entirely dependent on both pH and temperature of incubation. Low pH and high temperature were effective in accelerating the germination rate, the maximal germination level being reached at pH 4.0 and 60°C in the presence of 0.5 M NaNO₂. On the basis of germination rate, the activation energy (μ) for the nitrite-induced germination calculated was approximately 9.9 kcal/mol. Germination was greatly stimulated after pretreatment of spores with DTT at pH 10.5 to remove the coats. Furthermore, cortical fragments prepared from spores of the same organism were lysed not only by lysozyme but also by NaNO_2. Hexosamine-containing material was also solubilized by these reagents. However, nitrite, unlike lysozyme, released a considerable amount of free hexosamine as well. These results suggest that nitrite-induced germination may involve an interaction of sodium nitrite as nitrous acid with some component of the cortex. A possible mechanism of nitrite-induced germination is discussed.