Aim and Background- Riboflavin or vitamin B2 is essential for body and has the significant role in the macromolecules metabolism. Human can not  to synthesize vitamin B2 and must be acquire it from food-diet. Riboflavin biosynthetic pathway have been studied in Bacteria. The aim of this study was to isolation and identification of Bacillus subtilis as, riboflavin producing bacteria from different soil of Iran.

Materials and Methods- Soil samples from different areas of Iran was collected and was purplated. Then was prepared pure isolated of  bacteria and was identified by biochemical and morphological tests. DNA was extracted from isolated of B. subtilis, and using PCR for detection of ribC gene. 

Results- Biochemical tests and PCR  results showed, from 100 soil samples, 19 B. subtilis was isolated that was contained ribC gene and could growth in riboflavin-free medium.

Conclusion- B. subtilis was isolated easy from the soil and ribC gene can coded riboflavin in B. subtilis. Riboflavin in B. subtilis was biosynthesed by regulatory region in the up-strand of ribC gene operon, therefore can be produced economic riboflavin from this bacterium that is appropriate for environmental.