Enzymatic Characterization of an Amine Oxidase from Arthrobacter sp. Used to Measure Phosphatidylethanolamine
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概要
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Ethanolamine oxidase was screened with the aim of using it to establish a novel enzymatic phosphatidylethanolamine assay. Ethanolamine oxidase activity was detected in the crude extract of Arthrobacter sp., and the enzyme was purified more than 15-fold in three steps with a 54% yield. SDS–PAGE revealed the presence of only one band, which migrated, with an apparent molecular mass of 70 kDa. Biochemical characterization of the enzyme showed phenylethylamine to be the preferred substrate, with the highest kcat/Km value. The primary structure, determined by sequencing the cloned gene, showed a high degree of identity to Cu-containing phenylethylamine oxidase (64%). When heterologously overexpressed in Escherichia coli, the enzyme exhibited only a trace of amine oxidase activity, but high levels of activity emerged after exposure to Cu2+, as is typical of recombinant copper amine oxidases. Preliminary application of this enzyme coupled with phospholipase D for determination of phosphatidylethanolamine is also described. This is the first enzymatic method for the measurement of phosphatidylethanolamine.
- 2008-10-23
著者
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Tamura Tomohiro
Res. Inst. Of Genome-based Biofactory Aist Toyohira-ku Sapporo 062-8517 Jpn
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Tamura Tomohiro
Research Institute Of Genome-based Biofactory Aist:laboratory Of Molecular Environmental Microbiolog
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Yasuda Yuko
Asahi Kasei Pharma Corporation
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SAKASEGAWA Shin-ichi
Asahi Kasei Corporation
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Ota Hiroko
Asahi Kasei Pharma Corporation
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Imamura Shigeyuki
Asahi Kasei Pharma Corporation
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Yasuda Yuko
Asahi Kasei Pharma Corp. Shizuoka
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Sakasegawa Shin-ichi
Asahi Kasei Pharma Corporation
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SASANO Yoshie
Kyushu University, Division of Biological Science and Technology, Department of Health Science, Scho
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HOKAZONO Eisaku
Kyushu University, Division of Biological Science and Technology, Department of Health Science, Scho
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OSAWA Susumu
Kyushu University, Division of Biological Science and Technology, Department of Health Science, Scho
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Osawa Susumu
Kyushu University Division Of Biological Science And Technology Department Of Health Science School
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Sasano Yoshie
Kyushu University Division Of Biological Science And Technology Department Of Health Science School
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Hokazono Eisaku
Kyushu University Division Of Biological Science And Technology Department Of Health Science School
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Tamura Tomohiro
Research Institute Of Genome-based Biofactory National Institute Of Advanced Industrial Science And
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Hino Emisa
Asahi Kasei Pharma Corporation
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Tamezane Hideto
Kyushu University, Division of Biological Science and Technology, Department of Health Science, School of Medicine
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