Abstract

Brucella sp. Lumazine synthase, the enzyme involved in riboflavin biosynthesis composed of 10 identical subunits.  According to extended applications of LS, it is necessary to set up a high yield expression and purification method for this enzyme.in current study, Lumazine synthase primary structure was achieved from NCBI and it was expressed by pET28a in BL21 E. coli. The optimum concentrations of IPTG and Kanamycin was evaluated and applied for high yield expression of rBLS. For LPS removal and purification of protein, ammonium precipitation and ion exchange chromatography were performed and no background in ELISA (against Brucella) was observed. Purification of protein with DEAE sephadex resulted in a single band purified rBLS. The results of rBLS based ELISA in comparison with c-ELISA (SVANOVIR kit) indicates that lumazine synthase cannot completely and efficiently detect Brucella. The approach applied in this study can be used in generation a relatively pure rBLS as a valuable recombinant product in vaccine industries.