Engineering Towards Catalytic Use of Fungal Class-II Peroxidases for Dye-Decolorizing and Conversion of Lignin Model Compounds
| Autor: | Andrew T. Smith, Kristiina Hildén, Johanna Rytioja, Matti Wahlsten, Jaana Kuuskeri, Taina Lundell, Usenobong F. Ufot, Paula Nousiainen, Wendy A. Doyle, Elodie Bentley, Martin Hofrichter |
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| Přispěvatelé: | Department of Food and Nutrition, Fungal Co-life, Omics and Ecophysiology Research Group, Department of Chemistry, Cyanobacteria research, Fungal Genetics and Biotechnology |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Organizational Behavior and Human Resource Management Strategy and Management 116 Chemical sciences Pharmaceutical Science Mycology 7. Clean energy Catalysis 03 medical and health sciences chemistry.chemical_compound Fungal genetics Drug Discovery Botany Lignin 1183 Plant biology microbiology virology 219 Environmental biotechnology Marketing Pharmacology biology Chemistry 414 Agricultural biotechnology 030104 developmental biology Biochemistry biology.protein Fungal biotechnology Lignocellulose biodegradation 1182 Biochemistry cell and molecular biology Heterologous expression Peroxidase |
| Zdroj: | Current Biotechnology. 6:116-127 |
| ISSN: | 2211-5501 |
| DOI: | 10.2174/2211550105666160520120101 |
| Popis: | Background. Manganese peroxidases (MnP) and lignin peroxidases (LiP) are haem-including fungal secreted class-II peroxidases, which are interesting oxidoreductases in protein engineering aimed at design of biocatalysts for lignin and lignocellulose conversion, dye compound degradation, activation of aromatic compounds, and biofuel production. Objective. Recombinant short-type MnP (Pr-MnP3) of the white rot fungus Phlebia radiata, and its manganese-binding site (E40, E44, D186) directed variants were produced and characterized. To allow catalytic applications, enzymatic bleaching of Reactive Blue 5 and conversion of lignin-like compounds by engineered class-II peroxidases were explored. Method. Pr-MnP3 and its variants were expressed in Escherichia coli. The resultant body proteins were lysed, purified and refolded into haem-including enzymes in 6-7% protein recovery, and examined spectroscopically and kinetically. Results. Successful production of active enzymes was attained, with spectral characteristics of high-spin class-II peroxidases. Recombinant Pr-MnP3 demonstrated high affinity to Mn2+, which was noticeably affected by single (D186H/N) and double (E40H+E44H) mutations. Without addition of Mn2+, Pr-MnP3 was able to oxidize ABTS and decolorize Reactive Blue 5. Pc-LiPH8, its Trp-radical site variants, and engineered CiP-LiP demonstrated conversion of veratryl alcohol and dimeric non-phenolic lignin-model compounds (arylglycerol-β-aryl ethers) with production of veratraldehyde, which is evidence for cation radical formation with subsequent Cα-Cβ cleavage. Pc-LiPH8 and CiP variants were able to effectively oxidize and convert the phenolic dimer (guaiacylglycerol-β–guaiacyl ether). Conclusion. Our results demonstrate suitability of engineered MnP and LiP peroxidases for dye-decolorizing, and efficiency of LiP and its variants for activation and degradation of phenolic and non-phenolic lignin-like aryl ether-linked compounds. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
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