Construction of a novel twin-arginine translocation (Tat)-dependent type expression vector for secretory production of heterologous proteins in Corynebacterium glutamicum

•A novel E. coli-C. glutamicum shuttle expression vector pAU3 was constructed for the secretory production of heterologous proteins in Corynebacterium glutamicum.•The vector pAU3 harbors the strong promoter tac-M, the consensus RBS sequence, and the strong Tat-type cgR_0949 signal sequence in C. glutamicum.•The applicability of pAU3 was confirmed by the highly efficient expression and secretion of the CAT protein in C. glutamicum.

ABSTRACTCorynebacterium glutamicum is recognized as a favorable host for the secretory production of heterologous proteins. However, there are few secretion-type expression vectors available for protein production in C. glutamicum. In this study, we constructed a shuttle expression vector pAU3, which harbors the strong promoter tac-M for constitutive gene transcription, the consensus RBS sequence for protein translation, and the strong cgR_0949 signal sequence for protein secretion via the Tat pathway in C. glutamicum. The applicability of pAU3 was confirmed by the highly efficient expression and secretion of the CAT protein in C. glutamicum. The vector pAU3 is highly useful for secretory production of heterologous proteins in C. glutamicum.

Graphical abstractMap of secretion-type expression vector pAU3 (A) and its applicability for CAT protein secretion in C. glutamicum (B and C)Figure optionsDownload full-size imageDownload as PowerPoint slide