NDLI: Soluble expression and purification of bioactive interleukin 33 in E. coli

Content Provider	Springer Nature Link
Author	Do, Bich Hang Park, Sangsu Kwon, Grace G. Nguyen, Minh Tan Kang, Hyo Jeong Song, Jung A Yoo, Jiwon Nguyen, Anh Ngoc Jang, Jaepyeong Jang, Mihee Lee, Sunju So, Seoungjun Sim, Sungrak Jin, Jonghwa Lee, Kyung Jin Osborn, Mark J. Choe, Han
Copyright Year	2017
Abstract	Interleukin-33 (IL-33) is one of the important alarmins of the immune system and possesses dual functions as an anti- or pro-inflammatory molecule. The production of this cytokine in E. coli is hampered by the insoluble expression in the cytoplasm, resulting in inclusion body formation. In this study, the expression of IL-33 was optimized by fusing the N-terminus of IL-33 with several solubilizing tags that act as chaperones for proper protein folding: maltose binding protein (MBP), b´a´ domain of protein disulfide isomerase (PDIb´a´) and glutathione Stransferase (GST). The expression of the fusion proteins was stimulated by 0.5 mM IPTG at different temperatures, 37, 30, 25, and 18°C. As a result, IL-33 was expressed highly and in soluble form in the cytoplasm of E. coli when fused with MBP or PDIb´a´ tags in the presence of 0.5 mM IPTG at 25 or 30°C. We describe a simple purification procedure of IL-33 from the PDIb´a´-IL-33 construct using immobilized metal affinity chromatography (IMACs) with supplementary of tobacco etch virus (TEV) protease for tag removal. The high bioactivity of purified IL-33 on the proliferation and activation of macrophages was confirmed by MTT and nitrite releasing assays using RAW 264.7 These data show an improved method for producing high grade and yield IL-33.
Starting Page	256
Ending Page	264
Page Count	9
File Format	PDF
ISSN	12268372
Journal	Biotechnology and Bioprocess Engineering
Volume Number	22
Issue Number	3
e-ISSN	19763816
Language	English
Publisher	The Korean Society for Biotechnology and Bioengineering
Publisher Date	2017-07-19
Publisher Place	Seoul
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	human Interleukin 33 (hIL-33) E. coli soluble expression macrophage activation purification macrophage proliferation Biotechnology Industrial and Production Engineering
Content Type	Text
Resource Type	Article
Subject	Bioengineering Biomedical Engineering Applied Microbiology and Biotechnology Biotechnology

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