Production of functional proteins by using secretory signal peptide of marine bacteria origin

Abstract

We isolated chitosanase secreting B. subtilis CH2 (Oh et al., 2011) and identified the chitosanase sequence. Analyzed the sequence showed that it consisted of 813 bp, including 87 bp signal sequence. The signal sequence leads the target protein to the cell-membrane of the B. subtilis CH2 and then secret the chitosanase out of the cell. The chitosanase sequence including signal peptide was cloned into pET11a vector without fusion and expressed in E. coli BL21(DE3). The expressed chitosanase in E. coli showed two distinct bands which represent the pro-chitosanase in cytoplasm and mature chitosanase in periplasm. Time frame induction and results showed that muture chitosanase was increased. Subsequently, we linked chitosanase and xylanase signal sequence of B. subtilis CH2 in front of Human Epidermal Growth Factor (hEGF), Human Superoxide Dismutase (hSOD) and Human Peroxiredoxin 6 (hPRXD6). Then, it expressed in E. coli BL21(DE3) and analyzed by SDS-PAGE. These signal peptides are useful for production of biomedicine in E. coli.tein to the cell-membrane of the B. subtilis CH2 and then secret the chitosanase out of the cell. The chitosanase sequence including signal peptide was cloned into pET11a vector without fusion and expressed in E. coli BL21(DE3). The expressed chitosanase in E. coli showed two distinct bands which represent the pro-chitosanase in cytoplasm and mature chitosanase in periplasm. Time frame induction and results showed that muture chitosanase was increased. Subsequently, we linked chitosanase and xylanase signal sequence of B. subtilis CH2 in front of Human Epidermal Growth Factor (hEGF), Human Superoxide Dismutase (hSOD) and Human Peroxiredoxin 6 (hPRXD6). Then, it expressed in E. coli BL21(DE3) and analyzed by SDS-PAGE. These signal peptides are useful for production of biomedicine in E. coli.