Purification and characterisation of new laccase from Trametes polyzona WRF03
| Autor: | Jerry Okwudili Udeh, Tobechukwu Christian Ezike, F. C. Chilaka, AL Ezugwu, S. O. O. Eze |
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| Rok vydání: | 2020 |
| Předmět: |
Laccase
Chromatography ABTS biology lcsh:Biotechnology Coomassie Brilliant Blue Oligonucleotide primers Trametes polyzona WRF03 Applied Microbiology and Biotechnology Enzyme assay chemistry.chemical_compound Dye decolourisation chemistry lcsh:TP248.13-248.65 biology.protein Sodium azide Malachite green Methylene blue Sodium cyanide ComputingMethodologies_COMPUTERGRAPHICS Research Article Biotechnology |
| Zdroj: | Biotechnology Reports Biotechnology Reports, Vol 28, Iss, Pp e00566-(2020) |
| ISSN: | 2215-017X |
| DOI: | 10.1016/j.btre.2020.e00566 |
| Popis: | Graphical abstract Highlights • Trametes polyzona WRF03 produced high yield of true laccase. • Trametes polyzona WRF03 laccase was relatively pH and temperature stable. • Fe2+, sodium azide and sodium cyanide greatly inhibited laccase activity. • Trametes polyzona WRF03 laccase decolorised many classes of synthetic dyes. The molecular screening for laccase specific gene sequences in Trametes polyzona WRF03 (TpWRF03) using designed oligonucleotide primers analogous to the conserved sequences on the copper-binding regions of known laccases showed positive amplification with an amplicon size corresponding to 1500 bp. The purified TpWRF03 laccase (TpL) is a monomer with a molecular weight corresponding to 66 kDa. The enzyme had an optimal pH of 4.5 and temperature of 55 °C. TpL was most stable within pH of 5.5–6.5 and at a temperature range of 40–50 °C. Sodium azide, sodium cyanide and Fe2+ greatly inhibited the enzyme activity. TpL showed more than 50 % decolourisation efficiency on coomassie brilliant blue (72.35 %) and malachite green (57.84 %) but displayed low decolourisation efficiency towards Azure B (1.78 %) and methylene blue (0.38 %). The results showed that TpWRF03 produces high-yield of true laccase with robust properties for biotechnological applications. |
| Databáze: | OpenAIRE |
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