African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12513

Full Length Research Paper

Screening for the optimal induction parameters for periplasmic producing interferon-α2b in Escherichia coli

Siti Nor Ani Azaman1, Ramakrishnan Nagasundara Ramanan1, Joo Shun Tan1, Raha Abdul Rahim2, Mohd Puad Abdullah2 and Arbakariya B. Ariff1, 3*
1Institute of Bioscience, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia. 2Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor , Malaysia. 3Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia.
Email: [email protected]

  •  Accepted: 05 August 2010
  •  Published: 20 September 2010

Abstract

Screening for optimum induction parameters to improve the production of periplasmic interferon-α2b (PrIFN-α2b) by recombinant Escherichia coli was conducted using shake flask culture. Recombinant E. coli Rosetta-gami 2(DE3) harboring the plasmid pET26b containing IFN-α2b gene under the control of the T7lac promoter was used, where the induction was accomplished by isopropyl β-D-1-thiogalactopyranoside (IPTG). The induction parameters (inducer concentration, point of induction, induction temperature and the length of induction) were analyzed to find the suitable range to be used for further optimization process. From the analysis, narrow range of induction temperature from 16 to 30°C and IPTG lower than 2 mM were found suitable for induction of PrIFN-α2b. On the other hand, early log phase was the preferred time to initiate the induction and the length of induction was dependent on the combination of other induction parameters used. 

 

Key words: Interferon-α2b (IFN-α2b), induction parameter, Escherichia coli,periplasm, shake flask culture.

Abbreviation

PrIFN-α2b, Periplasmic interferon-α2b; IPTG, isopropyl β-D-1-thiogalactopyranoside.