caa a22 4500 605498318 CHVBK 20210128100545.0 cr unu---uuuuu 210128e20150801xx s 000 0 eng 10.1007/s00253-015-6441-4 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6441-4 Soluble expression, rapid purification, and characterization of human interleukin-24 (IL-24) using a MBP-SUMO dual fusion system in Escherichia coli [Elektronische Daten] [Jian Zhang, Xinxin Lv, Rui Xu, Xinyi Tao, Yuguo Dong, Aiyou Sun, Dongzhi Wei] Interleukin-24 (IL-24), a cytokine belonging to the IL-10 family, can selectively induce apoptosis in a broad range of tumor cells without harming normal cells. The efficient and soluble expression of bioactive recombinant IL-24 in Escherichia coli remains an obstacle because of aggregation and insufficient yield. In this study, a fusion of the small ubiquitin-related modifier (SUMO) or maltose-binding protein (MBP) has shown potential in facilitating the produce of IL-24. Thus, a new construct for MBP-SUMO-IL-24 expression would be a promising approach. Our results showed that the MBP-SUMO-IL-24 fusion protein was efficiently expressed as a soluble protein. SUMO protease-mediated cleavage at the SUMO/IL-24 junction released the recombinant IL-24 from the fusion protein. In addition, a His6 tag fused upstream of SUMO allowed for one-step purification through nickel affinity chromatography. Cleavage of the MBP-SUMO tag on the column resulted in the release of purified IL-24 and simplified the purification process. The final yield of IL-24 with approximately 90% purity was 19mg/L in flask fermentation. In vitro activity assays demonstrated that the purified IL-24 could induce apoptosis in MCF-7 breast cancer cells, but not normal NHLF cells, in a dose-dependent manner. In summary, we developed a novel method to express soluble and bioactive IL-24 protein in prokaryotic cells. Springer-Verlag Berlin Heidelberg, 2015 Interleukin-24 (IL-24) nationallicence Small ubiquitin-related modifier (SUMO) nationallicence Maltose-binding protein (MBP) nationallicence Soluble expression nationallicence One-step purification nationallicence Apoptosis nationallicence Zhang Jian State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 130 Meilong Road, 200237, Shanghai, People's Republic of China aut Lv Xinxin State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 130 Meilong Road, 200237, Shanghai, People's Republic of China aut Xu Rui State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 130 Meilong Road, 200237, Shanghai, People's Republic of China aut Tao Xinyi State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 130 Meilong Road, 200237, Shanghai, People's Republic of China aut Dong Yuguo State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 130 Meilong Road, 200237, Shanghai, People's Republic of China aut Sun Aiyou State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 130 Meilong Road, 200237, Shanghai, People's Republic of China aut Wei Dongzhi State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 130 Meilong Road, 200237, Shanghai, People's Republic of China aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/16(2015-08-01), 6705-6713 0175-7598 99:16<6705 2015 99 253 https://doi.org/10.1007/s00253-015-6441-4 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6441-4 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Zhang Jian State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 130 Meilong Road, 200237, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Lv Xinxin State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 130 Meilong Road, 200237, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Xu Rui State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 130 Meilong Road, 200237, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Tao Xinyi State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 130 Meilong Road, 200237, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Dong Yuguo State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 130 Meilong Road, 200237, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Sun Aiyou State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 130 Meilong Road, 200237, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Wei Dongzhi State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 130 Meilong Road, 200237, Shanghai, People's Republic of China aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/16(2015-08-01), 6705-6713 0175-7598 99:16<6705 2015 99 253