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Immobilized Zinc Affinity Chromatography of Pectin Hydroxamic Acids for Purification of Trypsin Inhibitors from Soybean and Sweet Potato
- Source :
- Journal of Agricultural and Food Chemistry. 53:10219-10223
- Publication Year :
- 2005
- Publisher :
- American Chemical Society (ACS), 2005.
-
Abstract
- Commercial pectin (with a 94% degree of esterification, DE94) suspended in methanol was reacted with methanolic alkaline hydroxylamine at room temperature for 20 h to prepare pectin hydroxamic acids (PHAs). The prepared PHA was coupled to the epoxy-activated Sepharose 6B gel to get immobilized PHA resins. The immobilized PHA resin was then balanced in column with 2 mM ZnCl2 in 50 mM Tris-HCl buffer (pH 7.9) to test the immobilized Zn-PHA gel as solid phase for immobilized metal affinity chromatography for the purification of trypsin inhibitors (TIs) from soybean and sweet potato. Using TI activity staining, it was found that purified TIs from the commercial soybean and sweet potato after trypsin affinity column purification could be adsorbed onto an immobilized Zn-PHA affinity column and eluted by 100 mM EDTA in 10 mM Tris-HCl buffer (pH 7.9). The immobilized Zn-PHA affinity column was used for TI purifications from crude extracts of sweet potato. The recovery of TI activity for one step was 90%, with 19.74-fold purification increase.
- Subjects :
- food.ingredient
Pectin
chemistry.chemical_element
Zinc
Hydroxamic Acids
Polysaccharide
Chromatography, Affinity
chemistry.chemical_compound
Hydroxylamine
food
Affinity chromatography
medicine
Ipomoea batatas
Phytohemagglutinins
chemistry.chemical_classification
Hydroxamic acid
Chromatography
Sepharose
General Chemistry
Trypsin
chemistry
Pectins
Electrophoresis, Polyacrylamide Gel
Soybeans
Methanol
Trypsin Inhibitors
General Agricultural and Biological Sciences
medicine.drug
Subjects
Details
- ISSN :
- 15205118 and 00218561
- Volume :
- 53
- Database :
- OpenAIRE
- Journal :
- Journal of Agricultural and Food Chemistry
- Accession number :
- edsair.doi.dedup.....49f9293d96a2743db46ca6d35b3a8356