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Optimization of Operating Conditions of Immobilized Penicillin G Acylase (PGA) Enzyme for Amoxicillin Synthesis

Darvish, Mahdi | 2009

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  1. Type of Document: M.Sc. Thesis
  2. Language: Farsi
  3. Document No: 39221 (06)
  4. University: Sharif University of Technology
  5. Department: Chemical and Petroleum Engineering
  6. Advisor(s): Roosta Azad, Reza; Aalemzadeh, Iran
  7. Abstract:
  8. Penicillin G Acylase is the key enzyme in producing semi-synthetic antibiotics such as Ampicillin and Amoxicillin. In this study, we used E.coli ATCC 11105 in order to produce it. This enzyme was an intracellular one which was produced under suitable conditions. Crude enzyme was released after cell disruption by ultrasonic waves (sonication method) and was partially purified by ammonium sulphate precipitation. Immobilization is investigated at different pHs around the iso-electric pH of Penicillin G Asylase enzyme. Therefore, optimum pH and optimum concentration of ammonium sulphate cuts were determined. Also immobilization process of this enzyme was studied on chitosan polymer, activated by glutaraldehyde, using covalent method in order to produce cheap biocatalyst. We used statistical methods in order to survey the effect of every influential parameter on immobilization and their interactions. Response Surface Methodology helps us optimizing effect of various variables with minimum number of experiments. By this method, values of glutaraldehyde concentration, enzyme load, and pH in this enzyme immobilization process were investigated. The results showed that the enzyme had its maximum activity in 2.7% glutaraldehyde concentration, 7.18 mg protein/gr polymer, and pH=8.15, which is 1.55 U/gr polymer
  9. Keywords:
  10. Immobilization ; Chitosan ; Response Surface Methodology ; Purification ; Penicillin G Acylase Enzyme ; Aluminum Sulphate

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