Bacillus subtilis an isolate of bacillus genus was obtained from the laboratories of Ministry of Science and Technology. The best efficient Bacillus subtilis isolate in cellulose and semi-cellulose hydrolysis was treated with Dielectric-barrier discharge (DBD)- Atmospheric cold plasma technique (non-thermal) using atmospheric air by exposing them at different times (2, 3, 4 and 5) min separately as a first stage and (60) seconds after any treatment separately as second stage (after 48 hour) in distance between the plasma source and the sample was fixed at 0.5 cm. The results showed a variation in the growth of the isolate according to the exposure time by the appearance of culture turbidity and the estimation of the optical density, where positive results appear between exposure times, the amount of optical density, and the cellulose and semi-cellulose decomposition into glucose. Bacillus subtilis increased its efficacy in cellulosic hydrolysis and semi-cellulosic materials, Bacillus subtilis showed malleability and ability to increase the efficiency in cellulose and semi-cellulose materials hydrolysis. We conduct a new and extensive study by using cold plasma technique to increases the hydrolysis efficacy of food microorganisms.Bacillus subtilis an isolate of bacillus genus was obtained from the laboratories of Ministry of Science and Technology. The best efficient Bacillus subtilis isolate in cellulose and semi-cellulose hydrolysis was treated with Dielectric-barrier discharge (DBD)- Atmospheric cold plasma technique (non-thermal) using atmospheric air by exposing them at different times (2, 3, 4 and 5) min separately as a first stage and (60) seconds after any treatment separately as second stage (after 48 hour) in distance between the plasma source and the sample was fixed at 0.5 cm. The results showed a variation in the growth of the isolate according to the exposure time by the appearance of culture turbidity and the estimation of the optical density, where positive results appear between exposure times, the amount of optical density, and the cellulose and semi-cellulose decomposition into glucose. Bacillus subtilis increased its efficacy in cellulosic hydrolysis and semi-cellulosic materials, Bacillus subtilis showed malleability and ability to increase the efficiency in cellulose and semi-cellulose materials hydrolysis. We conduct a new and extensive study by using cold plasma technique to increases the hydrolysis efficacy of food microorganisms.