Expression and secretion of heterologous proteases by Corynebacterium glutamicum

Abstract

Genes encoding the basic protease of Dichelobacter nodosus (bprV) and the subtilisin of Bacillus subtilis (aprE) were cloned and expressed in Corynebacterium glutamicum. In each case, enzymatically active protein was detected in the supernatants of liquid cultures. While the secretion of subtilisin was directed by its own signal peptide, the natural signal peptide of the bprV basic protease did not facilitate secretion. A hybrid aprE-bprV gene in which the promoter and signal peptide coding sequences of subtilisin replaced those of bprV could be expressed, and basic protease was secreted by C. glutamicum. Expression of these proteases in C. glutamicum provides an opportunity to compare protein secretion from this gram-positive host with that from other gram-positive and gram-negative bacteria.

Proteomics of corynebacteria: From biotechnology workhorses to pathogens

“…While the Sec pathway translocates unfolded proteins across the cytoplasmic membrane, the twin‐arginine translocation pathway primarily facilitates the passage of folded proteins, a feature that might be beneficial for biotechnological purposes 56. Furthermore, the potential of C. glutamicum for industry‐scale protein production was investigated in several studies since C. glutamicum culture supernatants exhibit no protease activity 57 and compared to bacilli only small amounts of extracellular proteins are secreted by this bacterium 54, 58–63. In summary, the results obtained by several studies indicate that C. glutamicum might be used as a general host for the production of proteins.…”

Plasmids of corynebacteria

“…With this system they were able to overexpress the protein, which was also secreted. Several other heterologous proteins such as ovine Q-interferon and proteases have been expressed successfully in C. glutamicum [46,47]. For the detection and analysis of promoter elements in corynebacteria several promoter probe vectors have been constructed.…”

Isolating promoters from Corynebacterium ammoniagenes ATCC 6871 and application in CoA synthesis

“…ammoniagenes native promoters to express foreign proteins in C. ammoniagenes [11][12][13]. Although some of these promoters are active in C. ammoniagenes, the activities are quite low [14].…”