Advancement in technology is continued as the time is passed. The biological active ingredients technology is the challenging issues for both the researchers and manufacturers. In last decade, much research has been happened about the potential health benefits of pro and prebiotic ingredients. Probiotics are the live microorganisms when given in adequate amount confer health benefits but the prebiotics are indigestible ingredients that enhance the activity of one or more probiotics in the colon. The therapeutic potential of these two biological active ingredients is the outcome of emerging field of biotechnology. A number of studies indicating therapeutic potential of probiotics regarding gastrointestinal health, cancer, constipation, immunomodulation, Helicobacter pylori, liver disease and urogenital infection have been documented. Similarly, prebiotics also improved helpful for many communicable and non-communicable diseases. It can be seen that advancement in the biotechnology field offer great choice to deliver a marvelous health advantages. Although the market for probiotic and prebiotic food and supplements has been increased in this new era but still need to develop food and supplements containing these two active ingredients with the cooperation of legal bodies of the country.

Withania coagulans is a shrub that has milk‐clotting activity. Its coagulation efficiency at different concentrations and incubation temperatures was measured for Mozzarella cheese from curd formation to its application on the pizza top. Curd made at higher W. coagulans concentration (1%) has less coagulation time (1272.99 ± 96.68 s), viscosity (148.58 ± 9.25 Pa s), pH (5.89 ± 0.05), and yield (7.87 ± 0.53%) as compared to rest of samples. Increasing temperature (39°C) also decreased coagulation time (1599.19 ± 380.06 s), viscosity (146.56 ± 15.51 Pa s), pH (6.01 ± 0.17), and yield (8.87 ± 0.48%) of curd. W. coagulans at 0.2% concentration showed caseinolytic activity (78.83 ± 1.97%) very close to rennet (84.12 ± 2.3%). High W. coagulans and temperature conditions lead to the grainy texture that results in the loss of proteins, fats, and moisture from the cheese. Highest protein (21.89 ± 0.8%; 23.39 ± 2.15%), fat (17.64 ± 0.62%; 18.50 ± 0.32%), and moisture (43.55 ± 8.01%; 50.00 ± 6.13%) were observed in Mozzarella cheese at 0.2% W. coagulans concentration and 33°C temperature, respectively. This study showed that 0.2% extract of W. coagulans can be used to clot the milk for cheese production at 33°C. Novelty impact statement The interest in plant‐based coagulant consumption has been growing because of the rennet shortage and religious matters of some vegetarian people. A plant‐based coagulant from Withania coagulans has been reported for its milk‐clotting activity, but its behavior for Mozzarella cheese production from curd formation to its application on pizza topping needs detailed study for its use on an industrial scale. In this study, W. coagulans behavior was studied during curd formation. Its caseinolytic and proteolytic activities in comparison to rennet were studied. Mozzarella cheese physicochemical and functional characteristics on pizza topping were also described in this research.