The present study ofCassia obtusifolia leaves (Kawal) were obtained from Darfur region western Sudan (Kutum, Jineena and Zalingie). The objective of this study is to isolate bacteria from different samples of dried fermented cassia obtusifolia leaves (kawal) and to identify bacteria by using different biochemical tests,which is present in natural environment and fermented traditionally as survival, the study was carried out in Khartoum state-Sudan , during February 2019 to September 2019, the samples were collected from three area (Kutum, Jineena and Zalingie). The traditional method for fermentation was used and the bacteria were isolated by using nutrient agar. The biochemical tests were used to determine type of bacteria and the results were cited positive with catalase, citrate, urease, motility tests, while gas and indol were showed negative result, the microorganisms were examined ,.The sugar fermentation tests for bacteria were recorded positive with glucose, sucrose, mannitol tests, while it showed negative at lactose sugar. The results revealed that all the samples were positive in nutrient agar media and the isolation of microorganisms were showed that ,thebacillusubtilis bacteria is only microorganism that was determined in the fermented kawalleave.