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Separation and Purification of Industrial Proteins by Chromatography and Aqueous two Phase System

Yavari, Milad | 2011

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  1. Type of Document: M.Sc. Thesis
  2. Language: Farsi
  3. Document No: 41728 (06)
  4. University: Sharif University of Technology
  5. Department: Chemical and Petroleum Engineering
  6. Advisor(s): Vossoughi, Manouchehr; Seifkordi, Ali Akbar
  7. Abstract:
  8. Separation of biomolecules such as enzymes is the most important part of biotechnology process which 10-90 % of costs is for separation and purification. There are several methods for purification of bio materials and one of them is chromatography which use in enzyme purification but this method have several problem such as cost ,time consuming and scale up Because of this a suitable alternative demands for industrial separation. An alternative is aqueous two-phase systems (ATPS). In this project was studied about partial purification of alkaline protease produced by Bacillus licheniformis ATCC 21424 in aqueous two-phase system and comparison with chromatography method. an aqueous two-phase system (ATPS) of polyethylene glycol (PEG)-salt was used for partial purification of alkaline protease produced by Bacillus licheniformis ATCC 21424. The effect of PEG molecular weight (1500 ,10000), pH (7.5, 9, 10.5) and NaCl concentration as additive salt on partition coefficient, yield and purification factor (PF) for ATPS were studied. Cation and anion exchange chromatography were performed and the yield and PF were derived from experimental data. Although, the results of our study indicated that using ATPS would lead to lower PF than ion-exchange chromatography, but its simplicity and high yield in addition of lower investments would make ATPS as promising purification candidate in an industrial scale
  9. Keywords:
  10. Aqueous Two-Phase System ; Chromatography ; Alkaline Protease Enzyme ; Protein Separation ; Protein Purification

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