15139

13616

كارشناسي ارشد

بيوتكنولوژي

اصفهان : دانشگاه صنعتي اصفهان

1398

چهارده، 104ص.: مصور، جدول، نمودار.

آذر شاه پيري

اصغر طاهري كفراني، آقا فخر ميرلوحي

ابوذر سورني، امير مساح

1398/06/31

كتابنامه

مهندسي كشاورزي

مهندسي كشاورزي

1398/07/14

2561755

Heterolougous expression of Cu Zn superoxide dismutase from Avicennia marina in Escherichia coli Monireh Fesharaki Esfahani Monireh fesharaki@yahoo com July 2 2019 Department of Agricultural Biotechonolgy Isfahan University of Technology Isfahan 84156 83111 IranDegree M Sc Language FarsiSupervisor A Shahpiri a shahpiri@cc iut ac irAbstractSuperoxide dismutase SOD is an antioxidant enzyme which protects cells from destructiveeffects of superoxide anion This enzyme catalyze dismutation of anion superoxide into oxygenand hydrogen peroxide This unique enzyme is used in the treatment of diseases caused byoxidative stress It is also used as an anti inflammatory enzyme in medical science Sofar many efforts have been made to improve the efficiency and stability of this enzyme becauseof its importance in medical application The purpose of this research is cloning expression purification and characterization of a novel CuZn SOD from Avicennia marina AmCuZn SOD To this aim the coding sequence of CuZn SOD was cloned into expression vector pET 28a Then this recombinant plasmid was transformed to Escherichia coli strain Rosetta DE3 The protein was expressed as a His tag fusion protein after induction with IPTG in fourdifferent conditions in the medium with no metals in the presence of Cu2 Zn2 and both Cu2 and Zn2 The SOD in these three conditions was produced and purified by affinitychromatography The expressed protein contains 188 amino acid residues and its molecularweight is 19 kDa The activity of this enzyme was determined presence of Cu2 and or Zn2 aswell as control medium The stability of the enzyme was measured in the temperature range of25 75 C and pH range of 3 12 Base on the results we found that AmSOD has its best activityby addition of both copper and zinc ion to culture of bacteria The optimum temperature was25 C and the optimum pH was pH3 8 These results open new window for application ofrecombinant form of AmSOD in the medicine and industry Key words Antioxidant Enzyme Superoxide Dismutase Avicennia Marina Heterologous expression Activity Stability 107

آذر شاه پيري

اصغر طاهري كفراني، آقا فخر ميرلوحي

ابوذر سورني، امير مساح