Sweet Potato Acid Phosphatase Immobilized on Glutaraldehyde-Activated Aminopropyl Controlled-Pore Glass: Activation, Repeated Use and Enzyme Fatigue
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- Yamato Susumu
- Department of Analytical Chemistry, Faculty of Pharmaceutical Sciences, Niigata University of Pharmacy and Applied Life Sciences
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- Kawakami Nozomi
- Department of Analytical Chemistry, Faculty of Pharmaceutical Sciences, Niigata University of Pharmacy and Applied Life Sciences
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- Shimada Kenji
- Department of Analytical Chemistry, Faculty of Pharmaceutical Sciences, Niigata University of Pharmacy and Applied Life Sciences
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- Ono Masaki
- Analytical Laboratory, Self-Medication Laboratories, Taisho Pharmaceutical Co., Ltd.
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- Idei Naoko
- Analytical Laboratory, Self-Medication Laboratories, Taisho Pharmaceutical Co., Ltd.
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- Itoh Yuji
- Analytical Laboratory, Self-Medication Laboratories, Taisho Pharmaceutical Co., Ltd.
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- Tachikawa Eiichi
- Department of Pharmacology, School of Medicine, Iwate Medical University
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Abstract
Sweet potato acid phosphatase was covalently coupled with glutaraldehyde to aminopropyl controlled-pore glass, and used as a pre-column enzyme reactor. The immobilized enzyme reactor (IMER) was continuously operated using an automated chromatographic detection system we developed. Functional evaluation of the IMER was carried out by injecting ten samples on the same day at an injection amount of 1.25 nmol (62.5 nmol per ml) using riboflavin sodium phosphate (FMNs) as a substrate, and by prolonged use for ten months. The IMER exhibited decreased activity after repeated use for a total of 3000 samples, but about 75% of its original activity remained. The conversion rate of FMNs to riboflavin by IMER was increased from 89 to 97% by adding citrate, ethylenediaminetetraacetic acid disodium salt, etc., but especially by adding citrate. The increased conversion of FMNs to riboflavin due to the addition of citrate was probably not due to the chelation of heavy metal ions by citrate. We also investigated complex formation of acid phosphatase with the substrate FMNs using surface plasmon resonance to determine the effect of citrate on the processes of association and/or dissociation between the enzyme and substrate. Enzyme fatigue was also observed during the course of prolonged and repeated use.
Journal
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- Biological and Pharmaceutical Bulletin
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Biological and Pharmaceutical Bulletin 27 (2), 210-215, 2004
The Pharmaceutical Society of Japan
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Details 詳細情報について
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- CRID
- 1390001204624944512
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- NII Article ID
- 110003608731
- 50000391315
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- NII Book ID
- AA10885497
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- ISSN
- 13475215
- 09186158
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- NDL BIB ID
- 6827195
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- PubMed
- 14758035
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- Text Lang
- en
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- Data Source
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- JaLC
- NDL
- Crossref
- PubMed
- CiNii Articles
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- Abstract License Flag
- Disallowed