The Cytoplasmic and Periplasmic Expression Levels and Folding of Organophosphorus Hydrolase Enzyme in Escherichia coli
| Author | Ali Mohammad Latifi | en |
| Author | Khosro Khajeh | en |
| Author | Gholamreza Farnoosh | en |
| Author | Kazem Hassanpour | en |
| Author | Samaneh Khodi | en |
| Issued Date | 2015-12-01 | en |
| Abstract | Background: Organophosphorus hydrolase (OPH) is a type of organophosphate-degrading enzyme which is widely used in the bioremediation process. Objectives: In this study, the periplasmic and cytoplasmic productions and the activity of recombinant OPH in Escherichia coli were investigated and compared using two pET systems (pET21a and pET26b). Materials and Methods: The sequence encoding the opd gene was synthesized and expressed in the form of inclusion body using pET21a-opd and in the periplasmic space in pET26b-opd. Results: Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis showed a band of about 37 kDa with a maximum expression level at 30°C from pET21a-opd.However, the obtained results of the periplasmic space extraction of OPH (pET26b-opd) showed a very weak band, while the cytoplasmic expression of OPH (pET21a-opd) produced a strong protein band. Conclusions: The activities studied by the production of PNP were determined by following the increase at 410 nm. The maximum PNP was produced at 30°C with an optical density of 10.62 in the presence of cytoplasmic expression of OPH (pET21a-opd). Consequently, our results suggest cytoplasmic expression system as an appropriate candidate with a high amount of OPH in spite of inclusion body formation, which needs an additional refolding step. | en |
| DOI | https://doi.org/10.5812/jjm.17790 | en |
| Keyword | Organophosphorus | en |
| Keyword | Cytoplasmic | en |
| Keyword | Periplasmic | en |
| Keyword | <i>Escherichia coli</i> | en |
| Publisher | Brieflands | en |
| Title | The Cytoplasmic and Periplasmic Expression Levels and Folding of Organophosphorus Hydrolase Enzyme in Escherichia coli | en |
| Type | Research Article | en |
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