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studying the Effect of Adding Nanoparticles in Polymer/salt Aqueous Two Phase Systems for Separating of Alpha-amylase Enzyme

Dehnavi, Mohsen | 2016

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  1. Type of Document: Ph.D. Dissertation
  2. Language: Farsi
  3. Document No: 48662 (48)
  4. University: Sharif University of Technology
  5. Department: Institute for Nanoscience and Nanotechnology
  6. Advisor(s): Vossoughi, Manoochehr; Pazuki, Gholamreza
  7. Abstract:
  8. The effects of adding nanoparticles in partitioning of α-amylase in polymer/salt aqueous two phase system (ATPS) was studied in this research. First the binodal curves of magnesium sulfate salt with PEGs 2000,3000,4000,6000 and 10000 at 23 ° C has been plotted and the suitable system which was PEG 6000/ magnesium sulfate has been selected for studying the partitioning of enzyme. Thereupon the partition coefficient of α-amylase was measured and then effect of adding silica and graphene oxide nanoparticles was investigated. It was observed that nanoparticles caused a dramatic increase in partition coefficient and yield recovery (of top phase). For further investigation the partitioning of enzyme in magnesium sulfate/PEG 2000 and magnesium sulfate/PEG 4000 ATPSs was also studied and effect of Hyperbranched polymer and silica nanoparticles was investigated. Then with use of variety analysis methods such as CD, FTIR, SAXS, TEM, DLS, UV, PL mechanisms and causes of this increment has been studied. After that the structural changes of extracted enzyme have been investigated by using a variety of methods and finally the activity of enzyme was measured with use of satandard tests that showed extracted enzyme has retained 85% it’s native structure
  9. Keywords:
  10. Partitioning ; Silica Nanoparticles ; Aqueous Two-Phase System ; Alpha-amylase Enzyme ; Aqueous Two-Phase System ; Enzyme Structural Study

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