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Title page for etd-0626106-153200


URN etd-0626106-153200 Statistics This thesis had been viewed 3627 times. Download 1632 times.
Author Kuen-Shan Jaw
Author's Email Address ksjaw@cef.ntist.edu.tw
Department Bioengineering
Year 2005 Semester 2
Degree Ph.D. Type of Document Doctoral Dissertation
Language English Page Count 103
Title Purification of sporamin from sweet ptotao by aqueous two phase system and affinity chromatography
Keyword
  • trypsin inhibitor
  • affinity chromatography
  • aqueous two-phase system
  • aqueous two-phase system
  • affinity chromatography
  • trypsin inhibitor
  • Abstract In this study, two methods were used to the purification of Kunitz soybean trypsin inhibitor with aqueous two-phase system and affinity chromatography from sweet potato Tainong 57. In aqueous two-phase, the polyethylene glycol (PEG) and phosphate were used to construct an aqueous two-phase system (ATPS). Trypsin inhibitor was recovered with high yield and high concentration in the top PEG-rich phase when the aqueous two phase system was constructed using PEG 6000 (11%), phosphate (16%), KCl (9%) at pH 6.0. The purity of the trypsin inhibitor was enhanced at 3.7-fold, and the recovery was 95%. The purified trypsin inhibitor showed one visible band, and the molecular weight was 27 kDa by SDS-PAGE. In affinity chromatography, the immobilized trypsin on the glutaraldehyde activated chitosan beads was employed to purify trypsin inhibitor from the tuberous roots of sweet potato by means of affinity chromatography. Maximum trypsin activity (10 unit/g-beads) of immobilized beads was obtained in treating with 0.1 % (w/v) glutaraldehyde, adding trypsin solution (2 mg/mL, pH 8) and shaking at 37 ?C for 6 hours in this study. Fifty-six percent of the activity of the trypsin inhibitor in the crude extract of sweet potato could be recovered through affinity chromatography. Seven visible bands by SDS-PAGE were suspected to be trypsin inhibitors, and all of the molecular weights were more than 20 kDa. The trypsin inhibitor of sporamin could be rapidly purified by either aqueous two-phase systems or affinity chromatography from sweet potato Tainong 57.
    Advisor Committee
  • Duan, Kow Jen - advisor
  • Chen, C. Will - co-chair
  • none - co-chair
  • none - co-chair
  • none - co-chair
  • Sheu, Dey-Chyi - co-chair
  • Files indicate access worldwide
    Date of Defense 2006-05-10 Date of Submission 2006-06-26


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