Purification and Properties of a Phytase from Candida krusei WZ-001(ENZYMOLOGY, PROTEIN ENGINEERING, AND ENZYME TECHNOLOGY)
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概要
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A phytase from Candida krusei WZ-001 isolated from soil was purified to electrophoretic homogeneity by ion-exchange chromatography, hydrophobic interaction chromatography, and gel filtration. The phytase is composed of two different subunits with molecular masses of 116 kDa and 31 kDa on SDS-PAGE (or 120 kDa and 30 kDa on gel chromatography), with the larger subunit having a glycosylation rate of around 35%. The phytase has an optimum pH of 4.6, an optimum temperature of 40℃ and a pl value of 5.5. The phytase activity was stimulated by 2-mercapto-ethanol and dithiothreitol (DTT), and inhibited by Zn^2+, Mg^2+, iodoacetate, p-chloromercuriben-zoate (pCMB) and phenylmethylsulfonyl fluoride (PMSF). The phytase displayed a broad substrate specificity and the K_m for phytate was 0.03 mM. Phytate was sequentially hydrolyzed by the phytase. Furthermore, 1D and 2D NMR analyses and bioassay of myoinositol indicated that the end hydrolysis product of phytate was myoinositol 2-monophosphate.
- 社団法人日本生物工学会の論文
- 2002-11-25
著者
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Ohta Y
Graduate School Of Biosphere Sciences Hiroshima University
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FAN Sheng-Di
Department of Biological Engineering, Dalian Nationalities University, Economical and Technological
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Fan Sheng-di
Department Of Biochemical Engineering Dalian Nationalities University
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Fan Sheng-di
Department Of Bio-engineering Dalian University For Nationalities Economical And Technological Devel
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Quan Chun-shan
Graduate School Of Biosphere Sciences Hiroshima University:department Of Bio-engineering Dalian Univ
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Wang Yun-ji
Faculty Of Food Engineering Dalian Institute Of Light Industry
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ZHANG LING-HUA
Faculty of Food Engineering, Dalian Institute of Light Industry
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OHTA YOSHIYUKI
Graduate School of Biosphere Sciences, Hiroshima University
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Zhang Ling-hua
Faculty Of Food Engineering Dalian Institute Of Light Industry
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Ohta Yoshiyuki
Graduate School Of Biosphere Sciences Hiroshima University
関連論文
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- Intermolecular Communication on Lipid Bilayer Membrane. Control of Enzymatic Activity Triggered by a Lipid Signal
- Purification and Properties of a Phytase from Candida krusei WZ-001(ENZYMOLOGY, PROTEIN ENGINEERING, AND ENZYME TECHNOLOGY)
- Pathway of Dephosphorylation of myo-Inositol Hexakisphosphate by a Novel Phytase from Candida Krusei WZ-001
- Catalytic Functions of Artificial Enzyme Composed of Simple Vitamin B12 Model and Synthetic Bilayer Membrane.