RESEARCH ARTICLE


Enhanced Interferon-α2b Production in Periplasmic Space of Escherichia coli through Medium Optimization using Response Surface Method



Joo Shun Tan1, Ramakrishnan Nagasundara Ramanan1, Siti Nor Ani Azaman1, Tau Chuan Ling2, Mustapha Shuhaimi3, Arbakariya B. Ariff*, 1, 4
1 Unit of Immunotherapeutic & Vaccine, Laboratory of Molecular Biomedicine, Institute of Bioscience, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia
2 Department of Process and Food Engineering, Faculty of Engineering, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia
3 Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia
4 Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia


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Creative Commons License
© 2009 Tan et al.

open-access license: This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International Public License (CC-BY 4.0), a copy of which is available at: (https://creativecommons.org/licenses/by/4.0/legalcode). This license permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

* Address correspondence to this author at the Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia; Tel: +603 89467591; Fax: +603 89467510; E-mail: arbarif@biotech.upm.edu.my


Abstract

The influence of different carbon and nitrogen sources on growth of recombinant Escherichia coli and human interferon-α2b (IFN-α2b) production in periplasmic space was studied in shake flask culture. A statistical method based on Plackett-Burman design was used to screen the main medium components that greatly influenced the performance of the fermentation process. The optimization of medium was performed using response surface methodology (RSM) where three critical factors (glucose, yeast extract and peptone) were optimized using central composite design. The highest yield of periplasmic recombinant human interferon-α2b (PrIFN-α2b) (335.8 μg/L) was predicted to be obtained in optimized medium containing 5.47 g/L glucose, 55.24 g/L yeast extract and 42.27 g/L peptone.. The production of IFN-α2b in periplasmic space in optimized medium was about 2.5, 11.7 and 124.4 times higher than Terrific broth (TB), Luria-Bertani (LB), and minimal medium (M9), respectively.

Keywords: Escherichia coli, Human interferon-α2b, Plackett-Burman, Optimization, Response surface methodology.