Inactivation of Bacillus cereus endospores on black pepper by pulsed superheated steam system

In this study, a superheated steam (SHS) system was constructed to inactivate Bacillus cereus endospores on the surface of black pepper, and continuous and pulsed treatment was applied to compare sporicidal effects. Additionally, inactivation mechanisms were analyzed to investigate the differences between pulsed and continuous SHS treatments. SHS at 250 °C and 300 °C for 1 min achieved more than a 3 log reduction, whereas SHS at 200 °C for 1 min achieved less than 2 log reduction in the number of endospores. In addition, higher microbicidal effects were confirmed with pulsed SHS treatment with a shorter duty ratio. To elucidate the inactivation mechanisms, inner membrane damage (dipicolinic acid release), intracellular enzyme activities, and DNA integrity were measured after 300 °C SHS pulsed or continuous treatments. After pulsed SHS treatment for up to 20 s, intracellular enzymes were inactivated more rapidly than after continuous treatment, and more DPA was released after 40 s of treatment, indicating that enzyme inactivation occurred prior to inner membrane damage, and pulsed treatment accelerated this mode of action. DNA integrity was significantly lower after 60 s of pulsed or continuous treatment; however, there was no difference in between pulsed and continuous treatments. Our results provide fundamental insights for the sterilization of black pepper by SHS treatment in food industries.