A rapid purification method for soybean Bowman-Birk protease inhibitor using hydrophobic-interaction chromatography
| Autor: | Kyoko Udaka, Chikafusa Fukazawa, Nana Asare Yeboah, Masaomi Arahira |
|---|---|
| Rok vydání: | 1996 |
| Předmět: |
Chymotrypsin
Chromatography biology Chemistry Hydrophilic interaction chromatography Size-exclusion chromatography Extraction (chemistry) Molecular Sequence Data Caseins Trypsin Protease inhibitor (biology) biology.protein medicine Chromatography Gel Electrophoresis Polyacrylamide Gel Amino Acid Sequence Peptide sequence Ammonium sulfate precipitation Biotechnology medicine.drug Trypsin Inhibitor Bowman-Birk Soybean |
| Zdroj: | Protein expression and purification. 7(3) |
| ISSN: | 1046-5928 |
| Popis: | A protein fraction was isolated from defatted soybean flour by extraction at acid pH, 40% ammonium sulfate precipitation, hydrophobic interaction chromatography, and gel filtration chromatography. SDS-PAGE, under reducing conditions, confirmed it as a homogeneous preparation. This conclusion was consistent with N-terminal amino acid sequence data (20 cycles) which showed a major sequence identical to those reported for soybean Bowman-Birk-type protease inhibitor (BBI), and also indicated a minimum 95% purity based on recoveries of PTH-amino acid residues. The purified fraction inhibited both trypsin and chymotrypsin with average specific activities of 350 and 672 units mg(-1), respectively. Compared with classical BBI purification, this procedure is very rapid requiring only 72-96 h to achieve a yield of 37 mg purified BBI per 200 g starting material. |
| Databáze: | OpenAIRE |
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