Overexpression of Recombinant Human Beta Interferon (rhINF-β) in Periplasmic Space of Escherichia coli.
Human Interferon β (INF-β) is a member of cytokines family which different studies have shown its immunomodulatory and antiviral activities. In this study an expression vector was designed and constructed for expression of human INF-β-1b either in shake flasks or bench top bioreactor. The designed vector was constructed based upon pET-25b(+) with T7 promoter. Recombinant human beta interferon (rhINF-β) was codon optimized and overexpressed as a soluble, N-terminal pelB fusion protein and secreted into the periplasmic space of Escherichia coli BL21 (DE3). The sugar, Isopropyl-β-D-thiogalactopyranoside (IPTG) was used as a chemical inducer for rhINF-β production in the shake flasks and bench top bioreactor. Timing of beta interferon expression was controlled by using the T7 promoter. The rhINF-β protein was extracted from periplasmic space by osmotic shock treatment and the expression of the beta interferon encoding gene in random selected transformants, was confirmed by western and dot blot methods. The maximum of product formation achieved at the OD600nm = 3.42 was found to be 35 % of the total protein content of the strain which translates to 0.32 g L-1. The constructed vector could efficiently overexpress the rhINF-β into the periplasmic space of E. coli. The obtained yield of the produced rhINF-β was more than previous reports. The system is easily adapted to include other vectors, tags or fusions and therefore has the potential to be broadly applicable to express other recombinant proteins.
Department of Pharmaceutical Biotechnology, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Mohammad Hossein
MH
Morowvat
Biochemical Engineering Group, Biotechnology Research Center, Malek-Ashtar University of Technology, Tehran, Iran. ; Department of Bioscience Engineering, Faculty of New Sciences and Technologies, University of Tehran, Tehran, Iran.
Valiollah
V
Babaeipour
Center of Biotechnology and Life sciences, Shahid Chamran University of Ahvaz, Iran. ; Department of Bioscience Engineering, Faculty of New Sciences and Technologies, University of Tehran, Tehran, Iran.
Hamid
H
Rajabi-Memari
Department of Pharmaceutical Biotechnology, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Hossein
H
Vahidi
Neuroscience Research Center, Shahid Beheshti University of Medical Science, Tehran, Iran.
Nader
N
Maghsoudi
Escherichia coli; Expression system; Interferon beta; Periplasmic expression; Plasmid