title: Effect of periplasmic expression of recombinant mouse interleukin-4 on hydrogen peroxide concentration and catalase activity in Escherichia coli.
abstract: Oxidative stress occurs as a result of imbalance between generation and detoxification of reactive oxygen species (ROS). This kind of stress was rarely discussed in connection with foreign protein production in Escherichia coli. Relation between cytoplasmic recombinant protein expression with H(2)O(2) concentration and catalase activity variation was already reported. The periplasmic space of E. coli has different oxidative environment in relative to cytoplasm and there are some benefits in periplasmic expression of recombinant proteins. In this study, hydrogen peroxide concentration and catalase activity following periplasmic expression of mouse IL-4 were measured in E. coli. After construction of pET2mIL4 plasmid, the expression of recombinant mouse interleukin-4 (mIL-4) was confirmed. Then, the H(2)O(2) concentration and catalase activity variation in the cells were studied in exponential and stationary phases at various ODs and were compared to those of wild type cells and empty vector transformed cells. It was revealed that empty vector introduction and periplasmic recombinant protein expression increased significantly the H(2)O(2) concentration of the cells. However, the H(2)O(2) concentration in mIL-4 expressing cells was significantly higher than its concentration in empty vector transformed cells, demonstrating more effects of recombinant mIL-4 expression on H(2)O(2) elevation. Likewise, although catalase activity was reduced in foreign DNA introduced cells, it was more lowered following expression of recombinant proteins. Correlation between H(2)O(2) concentration elevation and catalase activity reduction with cell growth depletion is also demonstrated. It was also found that recombinant protein expression results in cell size increase.
authors.affiliation: Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Elnaz
authors.initials: E
authors.lastname: Mehdizadeh Aghdam
authors.forename: Lena
authors.initials: L
authors.lastname: Mahmoudi Azar
authors.forename: Abolfazl
authors.initials: A
authors.lastname: Barzegari
authors.forename: Farrokh
authors.initials: F
authors.lastname: Karimi
authors.forename: Majid
authors.initials: M
authors.lastname: Mesbahfar
authors.forename: Naser
authors.initials: N
authors.lastname: Samadi
authors.forename: Mohammad Saeid
authors.initials: MS
authors.lastname: Hejazi
mesh_terms: D000818:Animals; D001483:Base Sequence; D002374:Catalase / Q000378:metabolism*; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology / Q000254:growth & development / Q000378:metabolism*; D006861:Hydrogen Peroxide / Q000378:metabolism*; D015847:Interleukin-4 / Q000378:metabolism*; D051379:Mice; D019897:Periplasm / Q000378:metabolism*; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000378:metabolism
chemical_list: D017931:DNA Primers; D011994:Recombinant Proteins; D015847:Interleukin-4; D006861:Hydrogen Peroxide; D002374:Catalase