SUMO-fusion and autoinduction-based combinatorial approach for enhanced production of bioactive human interleukin-24 in
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BIOTECHNOLOGICALLY RELEVANT ENZYMES AND PROTEINS
SUMO-fusion and autoinduction-based combinatorial approach for enhanced production of bioactive human interleukin-24 in Escherichia coli Sana Tahir 1 & M. Mudassir Iqbal 1 & M. Waheed Akhtar 2 & Qingbing Wang 3 & Tao Sun 4 & Saima Sadaf 1 Received: 25 April 2020 / Revised: 15 September 2020 / Accepted: 21 September 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract High-level production of recombinant human interleukin-24 (IL-24), a multifunctional immunomodulatory cytokine, has been challenging due primarily to its aggregation as inclusion bodies in the bacterial host while persistent poor-expression in the insect/ mammalian expression systems. The present study presents a robust, vector-host combination (pE-SUMO-IL24), auto-inducible medium (YNG/M9NG), and a simple purification scheme for soluble, bioactive, and cost-effective production of native-like IL24 (nIL-24) in Escherichia coli. The final protein yield, following a three-step purification scheme (IMAC, SEC, dialysis), was 98 mg/L in shake-flask culture (with scale-up potential), which was several folds higher than reported earlier. In vitro cytotoxicity assays with HeLa and HCT116 cancer cell lines (performed using different concentrations of nIL-24) and the fluorescence activated cell sorting analysis (FACS) revealed a dose- and concentration-dependent increase in the population of pro-apoptotic cells with concomitant, statistically significant drop in the number of cells existent at Go/G1-, S-, and G2/M-phases (P < 0.002). The bioactive nIL-24, developed through this study, holds promise for use in further functional characterizations/applications. Key points • Yeast SUMO fusion partner at N-terminus for improved solubility of an otherwise insoluble IL-24 in E. coli. • Enhanced cell densities with concomitant several-fold increase in protein yield by lactose-inducible media. • Improved inhibition of cervical and colorectal carcinomas by native-like nIL-24 compared with Met-containing IL. • Heterologous nIL-24 may enable better understanding of the functional intricacies linked up with its unique cancer-specific features.
Keywords Auto-induction . Cancer cells’ inhibition . Flow cytometry . Immunomodulatory cytokine . Interleukin-24 . Small ubiquitin-like modifier protein (SUMO)
Sana Tahir and M. Mudassir Iqbal contributed equally to this work. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00253-020-10921-4) contains supplementary material, which is available to authorized users. * Saima Sadaf [email protected]
3
Department of Interventional Radiology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, 197 Rui Jin Er Road, Shanghai, People’s Republic of China
1
Biopharmaceuticals and Biomarkers Discovery Lab, Institute of Biochemistry and Biotechnology, University of the Punjab, Lahore 54590, Pakistan
4
2
School of Biological Sciences, University of the Punjab, Lahore 54590, Pakistan
Key Laboratory of Sma
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