High overexpression of dye decolorizing peroxidase TfuDyP leads to the incorporation of heme precursor protoporphyrin IX

Dana I. Colpa, Marco W. Fraaije*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

4 Citations (Scopus)

Abstract

Highlights

Dye decolorizing peroxidase TfuDyP binds heme and protoporphyrin IX in vivo.

The activity of TfuDyP is dependent on the expression level in E. coli.

Expression of fully functional DyPs can be tuned by the type of expression host and expression conditions.

The heterologous overexpression level of the bacterial dye decolorizing peroxidase TfuDyP in Escherichia coli was increased sixty fold to approximately 200 mg of purified enzyme per liter culture broth by fusing the enzyme to the small ubiquitin-related modifier protein (SUMO). The highly over expressed SUMO-TfuDyP was, however, almost inactive. Analysis of the enzyme by UV-vis absorption spectroscopy and high-resolution mass spectrometry showed that a large fraction of the highly over-expressed enzyme contained the iron deficient heme precursor protoporphyrin IX (PPIX) instead of heme. Here we show that the activity of the enzyme was dependent on the expression level of the protein. (C) 2016 Elsevier B.V. All rights reserved.

Original languageEnglish
Pages (from-to)372-377
Number of pages6
JournalJournal of Molecular Catalysis B: Enzymatic
Volume134
Issue numberPart B
DOIs
Publication statusPublished - Dec-2016
Event8th Meeting on OxiZymes - Wageningen, Netherlands
Duration: 3-Jul-20166-Jul-2016

Keywords

  • Dye decolorizing peroxidase
  • Thermobifida fusca
  • Protoporphyrin IX
  • THERMOBIFIDA FUSCA
  • ESCHERICHIA-COLI
  • PROTEINS
  • EXPRESSION
  • IRON
  • MONOOXYGENASE
  • HEMOPROTEINS
  • DISCOVERY
  • LIGNIN

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