Covalent Chromatography for Chymotrypsin-like Proteases Using a Diphenyl 1-Amino-2-phenylethylphosphonate Derivative
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- Ono Shin
- Graduate School of Science and Engineering, University of Toyama
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- Murai Junya
- Graduate School of Science and Engineering, University of Toyama
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- Furuta Seigo
- Graduate School of Science and Engineering, University of Toyama
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- Doike Kazuya
- Graduate School of Science and Engineering, University of Toyama
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- Manzaki Fumie
- Graduate School of Science and Engineering, University of Toyama
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- Yoshimura Toshiaki
- Graduate School of Science and Engineering, University of Toyama
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- Kuroda Hirofumi
- Department of Applied Chemistry and Chemical Engineering, Toyama National College of Technology
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- Umezaki Masahito
- Institute of Natural Medicine, University of Toyama
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- Oyama Hiroshi
- Faculty of Science and Engineering, Setsunan University
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Abstract
To establish a covalent chromatography system for purification of naturally occurring chymotrypsin-like serine proteases, a diphenyl 1-amino-2-phenylethylphosphonate derivative bearing Gly-Gly-Gly as a spacer was immobilized on the Sepharose 4FF gel. In this system, bovine chymotrypsin was selectively bound to the phosphonate immobilized on the gel and then released by the action of 2-pyridinealdoxime methiodide as the reactivated enzyme. Based on the study results for selective binding and reactivation conditions, chymotrypsin-like proteases from pancreatin (hog pancreas) were rapidly and highly purified within three hours.
Journal
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- Journal of Biological Macromolecules
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Journal of Biological Macromolecules 13 (3), 78-85, 2013
Japan Science Society of Biological Macromolecules
