Flagellin FliC of Salmonella Typhimurium is an important antigen which is able to stimulate immune response in both innate and adaptive immunes to help animals against the bacterium. Moreover, the antiflagellin antibody in vaccinated chickens was identified to be transmitted into eggs for protecting the newly hatched chickens from Salmonella. Thus, the gene coding for S. Typhimurium FliC was expressed in E. coli in previous study to investigate feasibility of the recombinant protein for Salmonella vaccine development. Protein FliC in this study was produced in fusion form with Thioredoxin thus it was designated TrxFliC. In TrxFliC production process, culture media took important role in cell-biomass of the recombinant E. coli BL21 as well as TrxFliC productivity. Among the investigated media: M9, M9Y, MYL and LB, the yield of TrxFliC in LB medium was much higher than in the others. Supplement of salts in M9 and glucose into LB medium resulted in nearly double of cell-biomass and significantly higher TrxFliC productivity than in LB medium. Further more, carbon sources also profoundly influenced on the growth and production of TrxFliC of the recombinant strain. Although glucose was identified as the most suitable carbon source for the E. coli strain growth, but was not good for resulting in high yield of TrxFliC. Using glycerol instead of glucose in LB medium supplemented with the salts of M9 resulted in lower specific growth rate of the recombinant strain (after 8 hours of cultivation, cell-biomass reached by the cell-biomass of the strain grown in medium containing glucose as carbon source at 6 hours) but TrxFliC productivity was much higher, approximately 3 folds. In the selected medium, 1.3 g TrxFliC from 11 culture was harvested.