Loading...

Simulation and Optimization of α-amylase Enzyme Purification Process
Using Column Chromatography

Rokhforouz, Mohammad-Reza | 2018

717 Viewed
  1. Type of Document: M.Sc. Thesis
  2. Language: Farsi
  3. Document No: 51549 (06)
  4. University: Sharif University of Technology
  5. Department: Chemical and Petroleum Engineering
  6. Advisor(s): Rosstaazad, Reza
  7. Abstract:
  8. Enzymes, which are biochemical catalysts, have important applications in different industries such as food, pharmaceutical, medical, chemical identification and etc. Several methods have been used for separation of enzymes among which precipitation, chromatography and two-phase system. In this work, purification of α-amylase has been done by Ammonium Sulfate and Acetone precipitation, gel filtration based on Sephadex, and PEG-salt aqueous two-phase system. Furthermore, mass and momentum conservations’ equations were solved simultaneously to simulate the process based on the finite element method (FEM). Crude enzyme was precipitated with different saturations of ammonium sulfate and the highest purification factor (PF) was obtained when the saturation was 60% (v/v). The obtained results from acetone precipitation revealed that using 50% acetone led to the lowest total activity and highest specific activity. To investigate the effect of enzyme to acetone ratio, different ratios of 1/6 up to 1/2 were considered. Highest PF, e.g., 3.11, was attained at the ratio of 1/2. Gel filtration with different lengths of 10, 20, 29 cm was prepared and it was found that increasing the length of the column results in higher PF values. Seven aqueous two-phase systems were made and the results showed that the mid-range value of PEG molecular weight gives the highest partition coefficient (Ke) and PF. Moreover, using KCl instead of NaCl leads to more efficient separation of proteins. Finally, a computational model was developed to gives new insights into the mass and momentum transport of solutes in the column chromatography
  9. Keywords:
  10. Alpha-amylase Enzyme ; Bacillus Subtilis ; Column Chromatography ; Aqueous Two-Phase System ; Simulation ; Purification

 Digital Object List

 Bookmark

No TOC