Expression and Purification of Nisin in Escherichia coli
Journal Title: SSR Institute of International Journal of Life Sciences (SSR-IIJLS) - Year 2018, Vol 4, Issue 4
Abstract
Fusion expression is a promising strategy for the production of bioactive peptides in Escherichia coli to enhance either soluble protein level or purification potential. Nisin is the bacteriocin that had been extensively studied and had been widely applied in many areas such as food and pharmaceutical. However, scientific reports on recombinant nisin production in E. coli are still insufficient. In this study, we constructed a new expression plasmid containing the coding sequence of NusA, hexahistidine and Lactobacillus lactic nisin coding sequence. Next, we introduced the expression plasmid into BL21 E. coli and produced the recombinant fusion nisin in E. coli. Recombinant E. coli extract was purified by nickel affinity chromatography and resulted in 77% yield with 55% purity. The bioactive nisin was successfully released from NusA 6xHis Nisin fusion protein by the endonuclease. The nisin showed its antibacterial activity on Listeria monocytogenes with an activity unit of 18.9 AU/mg. The nisin bioactivity has stable at the temperature range of 30–90oC and in pH range of 1–12. The results showed that the new construction was appropriate for production of nisin bioactive peptides.
Authors and Affiliations
Huynh Thi Xuan Mai, Nguyen Van Hau, Nguyen Hieu Nghia, Dang Thi Phuong Thao
Keywords
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- EP ID EP671026
- DOI 10.21276/ijlssr.2018.4.4.9
- Views 95
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