Simultaneous Determination of Manganese Peroxidase and Lignin Peroxidase by Capillary Electrophoresis Enzyme Assays

Sumire Kudo, Airi Harada, Hiroe Kubota, Keiko Sasaki, Takashi Kaneta

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Here, we developed an enzyme assay of manganese peroxidase (MnP) by capillary electrophoresis using an in-capillary reaction and applied it to a simultaneous assay of MnP and lignin peroxidase (LiP). The enzyme activity of MnP was determined from the peak area corresponding to Mn(III)-malonate produced by the plug-plug reaction between MnP and Mn(II) in a separation capillary. A background electrolyte containing 250 mM malonate buffer (pH 4.5) and 5 mM cetyltrimethylammonium bromide was employed for the separation of Mn(III)-malonate from MnP at -10 kV after a plug-plug reaction for 5 min. Although the assay permitted the determination of purified MnP, we found that both LiP and MnP have similar activities against their substrates, that is, LiP catalyzed the oxidation reaction of Mn(II) as well as MnP, whereas MnP catalyzed the oxidation reaction of veratryl alcohol which was the substrate used in the LiP assay developed previously. Thus, we proposed a method to discriminate MnP from LiP based on the difference in the activities of these enzymes to each substrate. Amounts of MnP and LiP in a mixture were successfully evaluated by the proposed method.

Original languageEnglish
Pages (from-to)7329-7333
Number of pages5
JournalACS Omega
Volume2
Issue number10
DOIs
Publication statusPublished - Jan 1 2017

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manganese peroxidase
Capillary electrophoresis
Lignin
Manganese
Assays
Enzymes
lignin peroxidase
Substrates
Oxidation

All Science Journal Classification (ASJC) codes

  • Chemical Engineering(all)
  • Chemistry(all)

Cite this

Simultaneous Determination of Manganese Peroxidase and Lignin Peroxidase by Capillary Electrophoresis Enzyme Assays. / Kudo, Sumire; Harada, Airi; Kubota, Hiroe; Sasaki, Keiko; Kaneta, Takashi.

In: ACS Omega, Vol. 2, No. 10, 01.01.2017, p. 7329-7333.

Research output: Contribution to journalArticle

Kudo, Sumire ; Harada, Airi ; Kubota, Hiroe ; Sasaki, Keiko ; Kaneta, Takashi. / Simultaneous Determination of Manganese Peroxidase and Lignin Peroxidase by Capillary Electrophoresis Enzyme Assays. In: ACS Omega. 2017 ; Vol. 2, No. 10. pp. 7329-7333.
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