Improved catalytic activities of a dye-decolorizing peroxidase (DyP) by overexpression of ALA and heme biosynthesis genes in Escherichia coli

Ramzi Ahmad Bazli, Jeong Eun Hyeon, Sung Ok Han

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Hemoproteins are biotechnologically important heme-containing enzymes, and their production often requires optimal supply of precursors, such as 5-aminolevulinic acid (ALA) and hemin. In this work, endogenous ALA biosynthesis in Escherichia coli was increased as a strategy to improve catalytic activities of a recombinant dye-decolorizing peroxidase (DyP) from Bacillus subtilis. The positive effects of this expression system (pHemAL-DyP) are compared with individually expressed DyP strains grown with (pDyP + Hemin) and without (pDyP) the addition of hemin. The pHemAL-DyP plasmid increased intracellular ALA up to 117.5 mg/L, a 4.0-fold increased from control strain (pET22b, 29.7 mg/L of ALA). Soret peak in the UV-vis spectra was the highest for pHemAL-DyP strain with Reinheitszahl (A408/A280) value of 0.65 indicating higher heme content in the DyP produced compared to pDyP (0.39) and pDyP + Hemin (0.46). Peroxidase activity was increased up to 66.7 U/mg in the pHemAL-DyP strain compared to 39.0 and 43.4 U/mg for pDyP and pDyP + Hemin, respectively. Decolorization percentage of Reactive Blue 19 dye was the highest in the pHemAL-DyP strain with 84.7% as compared to the pDyP (69.9%) and pDyP + Hemin (72.8%) systems. In brief, enzymatic properties of recombinant DyP were successfully enhanced using this genetic engineering strategy thus eliminating the need for costly exogenous precursors.

Original languageEnglish
Pages (from-to)1272-1276
Number of pages5
JournalProcess Biochemistry
Volume50
Issue number8
DOIs
Publication statusPublished - 27 Feb 2015
Externally publishedYes

Fingerprint

Aminolevulinic Acid
Biosynthesis
Heme
Escherichia coli
Peroxidase
Catalyst activity
Coloring Agents
Dyes
Genes
Hemin
Acids
Strain control
Genetic engineering
Genetic Engineering
Bacilli
Bacillus subtilis
Plasmids
Enzymes

Keywords

  • 5-Aminolevulinic acid
  • Dye-decolorization
  • Dye-decolorizing peroxidase
  • Heme peroxidase
  • Hemoprotein

ASJC Scopus subject areas

  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Bioengineering

Cite this

Improved catalytic activities of a dye-decolorizing peroxidase (DyP) by overexpression of ALA and heme biosynthesis genes in Escherichia coli. / Ahmad Bazli, Ramzi; Hyeon, Jeong Eun; Han, Sung Ok.

In: Process Biochemistry, Vol. 50, No. 8, 27.02.2015, p. 1272-1276.

Research output: Contribution to journalArticle

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