Sweet Potato Acid Phosphatase Immobilized on Glutaraldehyde-Activated Aminopropyl Controlled-Pore Glass : Activation, Repeated Use and Enzyme Fatigue(Miscellameous)
スポンサーリンク
概要
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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 1.25nmol (62.5nmol 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.
- 2004-02-01
著者
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Ono Mayumi
Department Of Pharmaceutical Oncology Graduate School Of Pharmaceutical Sciences Kyushu University
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Ono M
Departments Of Biochemistry Kyushu University School Of Medicine
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Itoh Yuji
Analytical Laboratory Self-medication Laboratories Taisho Pharmaceutical Co. Ltd.
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Ono M
Analytical Laboratory Self-medication Laboratories Taisho Pharmaceutical Co. Ltd.
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SHIMADA Kenji
Department of Analytical Chemistry, Faculty of Pharmaceutical Sciences, Niigata University of Pharma
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YAMATO Susumu
Department of Analytical Chemistry, Niigata College of Pharmacy
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TACHIKAWA Eiichi
Department of Pharmacology, School of Medicine, Iwate Medical University
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河上 奈保子
京都薬大 衛生化学
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KAWAKAMI Nozomi
Department of Pharmacology, Faculty of Pharmaceutical Sciences, The University of Tokushima
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Kawakami Naomi
Department Of Hospital Pharmacy Toyama Medical And Pharmaceutical University
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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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Idei Naoko
Analytical Laboratory Self-medication Laboratories Taisho Pharmaceutical Co. Ltd.
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Ono M
Faculty Of Pharmaceutical Sciences Tokushima Bunri University
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大和 進
Pharmaceutical And Analytical Chemistry Niigata Univ. Of Pharmacy And Applied Life Sciences
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Shimada Kenji
Department Of Analytical Chemistry Faculty Of Pharmaceutical Sciences Niigata University Of Pharmacy
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Tachikawa E
Department Of Pharmacology School Of Medicine Iwate Medical University
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Tachikawa Eiichi
Department Of Pharmacology Faculty Of Pharmacy Iwate Medical University
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Kawakami Nozomi
Department Of Pharmacology Faculty Of Pharmaceutical Sciences The University Of Tokushima
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Tachikawa Eiichi
Department Of Endocrine Pharmacology Tokyo University Of Pharmacy And Life Sciences
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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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