To increase the l-isoleucine production in Corynebacterium glutamicum by overexpressing the global regulator Lrp and the two-component export system BrnFE.The brnFE operon and the lrp gene were cloned into the shuttle vector pDXW-8 individually or in combination. The constructed plasmids were transformed into an l-isoleucine-producing strain C. glutamicum JHI3-156, and the l-isoleucine production in these different strains was analysed and compared. More l-isoleucine was produced when only Lrp was expressed than when only BrnFE was expressed. Significant increase in l-isoleucine production was observed when Lrp and BrnFE were expressed in combination. Compared to the control strain, l-isoleucine production in JHI3-156/pDXW-8-lrp-brnFE increased 63% in flask cultivation, and the specific yield of l-isoleucine increased 72% in fed-batch fermentation.Both Lrp and BrnFE are important to enhance the l-isoleucine production in C. glutamicum.The results provide useful information to enhance l-isoleucine or other branched-chain amino acid production in C. glutamicum.