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Expression and Characterization of a Dye-Decolorizing Peroxidase from Pseudomonas Fluorescens Pf0-1

Authors :
Nikola Lončar
Natalija Drašković
Nataša Božić
Elvira Romero
Stefan Simić
Igor Opsenica
Zoran Vujčić
Marco W. Fraaije
Source :
Catalysts, Vol 9, Iss 5, p 463 (2019)
Publication Year :
2019
Publisher :
MDPI AG, 2019.

Abstract

The consumption of dyes is increasing worldwide in line with the increase of population and demand for clothes and other colored products. However, the efficiency of dyeing processes is still poor and results in large amounts of colored effluents. It is desired to develop a portfolio of enzymes which can be used for the treatment of colored wastewaters. Herein, we used genome sequence information to discover a dye-decolorizing peroxidase (DyP) from Pseudomonas fluorescens Pf-01. Two genes putatively encoding for DyPs were identified in the respective genome and cloned for expression in Escherichia coli, of which one (PfDyP B2) could be overexpressed as a soluble protein. PfDyP B2 shows some typical features known for DyPs which includes the ability to convert dyes at the expense of hydrogen peroxide. Interestingly, t-butyl hydroperoxide could be used as an alternative substrate to hydrogen peroxide. Immobilization of PfDyP B2 in calcium-alginate beads resulted in a significant increase in stability: PfDyP B2 retains 80% of its initial activity after 2 h incubation at 50 °C, while the soluble enzyme is inactivated within minutes. PfDyP B2 was also tested with aniline and ethyl diazoacetate as substrates. Based on GC-MS analyses, 30% conversion of the starting material was achieved after 65 h at 30 °C. Importantly, this is the first report of a DyP-catalyzed insertion of a carbene into an N-H bond.

Details

Language :
English
ISSN :
20734344
Volume :
9
Issue :
5
Database :
Directory of Open Access Journals
Journal :
Catalysts
Publication Type :
Academic Journal
Accession number :
edsdoj.3997a5baf8244a7d9549ab88e9868bfd
Document Type :
article
Full Text :
https://doi.org/10.3390/catal9050463