Enzymatic characterization of an amine oxidase from arthrobacter sp. used to measure phosphatidylethanolamine

Hiroko Ota, Hideto Tamezane, Yoshie Sasano, Eisaku Hokazono, Yuko Yasuda, Shin Ichi Sakasegawa, Shigeyuki Imamura, Tomohiro Tamura, Susumu Osawa

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17 Citations (Scopus)

Abstract

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 Cu 2+, 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.

Original languageEnglish
Pages (from-to)2732-2738
Number of pages7
JournalBioscience, Biotechnology and Biochemistry
Volume72
Issue number10
DOIs
Publication statusPublished - Nov 5 2008

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

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    Ota, H., Tamezane, H., Sasano, Y., Hokazono, E., Yasuda, Y., Sakasegawa, S. I., Imamura, S., Tamura, T., & Osawa, S. (2008). Enzymatic characterization of an amine oxidase from arthrobacter sp. used to measure phosphatidylethanolamine. Bioscience, Biotechnology and Biochemistry, 72(10), 2732-2738. https://doi.org/10.1271/bbb.80365