Expression, characterization and 2,4,6-trichlorophenol degradation of laccase from Monilinia fructigena
| Autor: | Wen-Hua Bao, Yong Xue, Yong-Sheng Tian, Quan-Hong Yao, Zhen Zhang, Ri-He Peng, Gao Jianjie, Wei Zhao |
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| Rok vydání: | 2011 |
| Předmět: |
Genetic Vectors
Trichlorophenol Pichia pastoris Substrate Specificity chemistry.chemical_compound Ascomycota Enzyme Stability Genetics Benzothiazoles Isoelectric Point Molecular Biology Chromatography High Pressure Liquid Thermostability Laccase Ions Monilinia fructigena ABTS biology Guaiacol General Medicine biology.organism_classification Enzyme assay Recombinant Proteins Molecular Weight Kinetics Biodegradation Environmental chemistry Biochemistry biology.protein Electrophoresis Polyacrylamide Gel Indicators and Reagents Sulfonic Acids Copper Chlorophenols |
| Zdroj: | Molecular biology reports. 39(4) |
| ISSN: | 1573-4978 |
| Popis: | A novel laccase gene from Monilinia fructigena was synthesized chemically according to the yeast bias codon and integrated into the genome of Pichia pastoris GS115 by electroporation. The expressed enzyme was recovered from the culture supernatant and purified. The result of enzyme activity assay and SDS-PAGE demonstrated that the recombinant laccase was induced and extracellularly expressed in P. pastoris. Main biochemical properties of this laccase, such as thermodependence and thermostability, optimal pH and pH stability, and the effect of metal ions and inhibitors, were characterized. With 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonate (ABTS) as the substrate, MfLcc had its optimal pH at 3.5 and optimal temperature at 45°C. The Km values of the ABTS, guaiacol were 0.012 and 0.016 Mm, respectively, and the corresponding V (max) values are 243.9 and 10.55 Um min(-1) mg(-1), respectively. The recombinant laccase degraded 80% 2,4,6-trichlorophenol after 8 h under the optimal conditions. The recombinant strain and its laccase can be considered as candidate for treating waste water polluted with trichlorophenols. |
| Databáze: | OpenAIRE |
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