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- Type of Document: M.Sc. Thesis
- Language: Farsi
- Document No: 41118 (06)
- University: Sharif University of Technology
- Department: Chemical and Petroleum Engineering
- Advisor(s): Aalemzadeh, Iran; Kazemi, Akhtarolmolouk
- Abstract:
- In this study, Pseudomonas aeruginosa BBRC-10036 was used for lipase production. The organism secreted the enzyme extracellulary. First of all, effect of initial pH of the culture broth on lipase activity was studied in order to determine the optimum condition for lipase production. After production, this enzyme must be separated from culture and after that the enzyme must be purified for using in analysis and industry. Different methods are used for purification of the enzyme. In this research, first precipitation was used and then this lipase has been purified by Ion exchange Chromatography leading to 2.33- fold purification and 11.47% recovery. In precipitation by acetone, maximum recovery and purification factor were observed at 50% and in precipitation by ethanol, maximum yield and purification factor were observed at 60%. From among four organic solvent at 50% (v/v) (acetone, ethanol, methanol, n- propanol), maximum yield and purification factor were observed at 50% acetone. Then lipase from P.aeruginosa was entrapped onto calcium alginate beads and effect of independent variables such as alginate concentration (%w/v), CaCl2 concentration (M) and enzyme load (%v/v) on immobilization yield and activity of immobilized enzyme were investigated. Media optimization for production of lipase was carried out by Design- Expert 8 software. The optimum conditions were as follows: sodium alginate concentration 2.5% (w/v), calcium chloride concentration 2.5(M) and enzyme load 50% (v/v). At those conditions, the highest immobilization yield and the optimum activity of immobilized enzyme, respectively, obtained were 93.65% and 2.64 unit/gr(IME)
- Keywords:
- Pseudomonas Aeruginosa Bacteria ; Lipase Enzyme ; Production ; Immobilization ; Calcium Alginate ; Purification
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