title: Development of a secretion system for the production of heterologous proteins in Corynebacterium glutamicum using the Porin B signal peptide.
abstract: Corynebacterium glutamicum is one of the useful hosts for the secretory production of heterologous proteins because of intrinsic attributes such as the presence of few endogenous proteins and proteases in culture medium. Here, we report the development of a new secretory system for the production of heterologous proteins by using the porin B (PorB) signal peptide in C. glutamicum. We examined two different endoxylanases and an antibody fragment (scFv) as model proteins for secretory production. In the flask cultivations, all the examined proteins were successfully produced as active forms into the culture medium with high efficiency. For the high-level production of endoxylanase, fed-batch cultivation was also performed in a lab-scale (5L) bioreactor, and the endoxylanases were efficiently secreted in the culture medium at levels as high as 615mg/L. From the culture supernatant, the secreted endoxylanases could be purified with high purity via one-step affinity column chromatography.
authors.affiliation: Department of Chemical and Biomolecular Engineering, KAIST, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Seul Ji
authors.initials: SJ
authors.lastname: An
authors.forename: Sung Sun
authors.initials: SS
authors.lastname: Yim
authors.forename: Ki Jun
authors.initials: KJ
authors.lastname: Jeong
mesh_terms: D001407:Bacillus / Q000235:genetics; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D019149:Bioreactors; D003001:Cloning, Molecular; D048230:Corynebacterium glutamicum / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D043364:Endo-1,4-beta Xylanases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D018272:Porins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D021382:Protein Sorting Signals; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D057127:Single-Chain Antibodies / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D048372:Streptomyces coelicolor / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D018272:Porins; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; D043364:Endo-1,4-beta Xylanases