Effect of periplasmic expression of recombinant mouse interleukin-4 on hydrogen peroxide concentration and catalase activity in Escherichia coli

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• 作者：Elnaz Mehdizadeh Aghdam^a ; ¹ ; Lena Mahmoudi Azar^a ; ¹ ; Abolfazl Barzegari^b ; ¹ ; Farrokh Karimi^c ; Majid Mesbahfar^a ; Naser Samadi^d ; Mohammad Saeid Hejazi^a ; ^d ; ^{saeidhejazi@tbzmed.ac.ir} ; ^{msaeidhejazi@yahoo.com}
• 关键词：ROS ; reactive oxygen species ; mIL-4 ; mouse interleukin-4 ; MetE ; cobalamin-independent methionine synthase ; HPI ; hydroperoxidase I ; HPII ; hydroperoxidase II ; SOD ; superoxide dismutase ; OD ; optical density ; PCR ; polymerase chain reaction ; IPTG ; isopropyl-be
• 刊名：Gene
• 出版年：2012
• 出版时间：15 December, 2012
• 年：2012
• 卷：511
• 期：2
• 页码：455-460
• 全文大小：652 K

文摘

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 H₂O₂ 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 H₂O₂ 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 H₂O₂ concentration of the cells. However, the H₂O₂ 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 H₂O₂ elevation. Likewise, although catalase activity was reduced in foreign DNA introduced cells, it was more lowered following expression of recombinant proteins. Correlation between H₂O₂ 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.