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Enzyme and Microbial Technology
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Enzyme and Microbial Technology
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Saturation mutagenesis to improve the degradation of azo dyes by versatile peroxidase and application in form of VP-coated yeast cell walls

Authors: Karla Ilić Đurđić; Raluca Ostafe; Aleksandra Đurđević Đelmaš; Nikolina Popović; Stefan Schillberg; Rainer Fischer; Radivoje Prodanovic;

Saturation mutagenesis to improve the degradation of azo dyes by versatile peroxidase and application in form of VP-coated yeast cell walls

Abstract

Azo dyes are toxic and carcinogenic synthetic pigments that accumulate as pollutants in aquatic bodies near textile factories. The pigments are structurally diverse, and bioremediation is mostly limited to single dye compounds or related groups. Versatile peroxidase (VP) from Pleurotus eryngii is a heme-containing peroxidase with a broad substrate spectrum that can break down many structurally distinct pollutants, including azo dyes. The utilization of this enzyme could be facilitated by engineering to modify its catalytic activity and substrate range. We used saturation mutagenesis to alter two amino acids in the catalytic tryptophan environment of VP (V160 and A260). Library screening with three azo dyes revealed that these two positions had a significant influence on substrate specificity. We were able to isolate and sequence VP variants with up to 16-fold higher catalytic efficiency for different azo dyes. The same approach could be used to select for VP variants that catalyze the degradation of many other types of pollutants. To allow multiple cycles of dye degradation, we immobilized VP on the surface of yeast cells and used washed cell wall fragments after lysis. VP embedded in the cell wall retained ∼70 % of its initial activity after 10 cycles of dye degradation each lasting 12 h, making this platform ideal for the bioremediation of environments contaminated with azo dyes.

Peer-reviewed manuscript: [http://cherry.chem.bg.ac.rs/handle/123456789/3835]

Supplementary material: [http://cherry.chem.bg.ac.rs/handle/123456789/3836]

Countries
Germany, Serbia
Subjects by Vocabulary

Microsoft Academic Graph classification: Lysis Bioremediation Saturated mutagenesis Versatile peroxidase biology Chemistry Substrate (chemistry) Protein engineering Combinatorial chemistry Yeast biology.protein Peroxidase

Keywords

Bioengineering, Saccharomyces cerevisiae, Applied Microbiology and Biotechnology, Biochemistry, Pleurotus eryngii, Substrate Specificity, Cell Wall, Coloring Agents, Dye degradation, Biodegradation, Environmental, Peroxidases, Biocatalysis, Mutagenesis, Site-Directed, Protein engineering, Cell wall fragments, Yeast surface display, Oxidoreductases, Azo Compounds, Oxidation-Reduction, Biotechnology

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