Improved catalytic activities of a dye-decolorizing peroxidase (DyP) by overexpression of ALA and heme biosynthesis genes in Escherichia coli
- Authors
- Ramzi, Ahmad Bazli; Hyeon, Jeong Eun; Han, Sung Ok
- Issue Date
- 8월-2015
- Publisher
- ELSEVIER SCI LTD
- Keywords
- Dye-decolorizing peroxidase; Dye-decolorization; 5-Aminolevulinic acid; Heme peroxidase; Hemoprotein
- Citation
- PROCESS BIOCHEMISTRY, v.50, no.8, pp.1272 - 1276
- Indexed
- SCIE
SCOPUS
- Journal Title
- PROCESS BIOCHEMISTRY
- Volume
- 50
- Number
- 8
- Start Page
- 1272
- End Page
- 1276
- URI
- https://scholar.korea.ac.kr/handle/2021.sw.korea/92925
- DOI
- 10.1016/j.procbio.2015.05.004
- ISSN
- 1359-5113
- 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 coil 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 (A(408)/A(280)) 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. (C) 2015 Elsevier Ltd. All rights reserved.
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