Use of Bacteriocin Release Protein in E. Coli for Excretion of Human Growth Hormone into the Culture Medium

摘要: Because of the outer membrane barrier, gram–negative bacterium E. coli is generally not considered to be an appropriate host for excretion proteins into culture medium. However, we have been able efficiently excrete a mammalian protein, human growth hormone (hGH), medium using cells harboring two vectors, hGH secretion vector pOmpA–hGH2 and pCloDF 13–derived bacteriocin release protein (BRP) expression pJL3. Both vectors contain, in tandem, lpp lac promoter–operator system. The tandem promoter drives BRP both regulated by represser. In presence isopropyl–1–thio–β–D–galactopy ranoside (IPTG) at low concentration (20 μM), OmpA–hGH hybrid were expressed mature was released excreted contained correct amino terminus corresponding derived from pituitary gland, indicating processing OmpA signal peptide. efficient it purified greater than 98% purity single step reversed–phase column chromatography.