Ali, E., Abd ellatif, S., Abdel Razik, E. (2020). Production, Purification, Characterization and Immobilization of Laccase from Phoma betae and its Application in Synthetic Dyes Decolorization. Egyptian Journal of Botany, 60(1), 301-312. doi: 10.21608/ejbo.2019.19226.1381
Eman Abdullah M. Ali; Sawsan Abd ellatif; Elsayed Abdel Razik. "Production, Purification, Characterization and Immobilization of Laccase from Phoma betae and its Application in Synthetic Dyes Decolorization". Egyptian Journal of Botany, 60, 1, 2020, 301-312. doi: 10.21608/ejbo.2019.19226.1381
Ali, E., Abd ellatif, S., Abdel Razik, E. (2020). 'Production, Purification, Characterization and Immobilization of Laccase from Phoma betae and its Application in Synthetic Dyes Decolorization', Egyptian Journal of Botany, 60(1), pp. 301-312. doi: 10.21608/ejbo.2019.19226.1381
Ali, E., Abd ellatif, S., Abdel Razik, E. Production, Purification, Characterization and Immobilization of Laccase from Phoma betae and its Application in Synthetic Dyes Decolorization. Egyptian Journal of Botany, 2020; 60(1): 301-312. doi: 10.21608/ejbo.2019.19226.1381
Production, Purification, Characterization and Immobilization of Laccase from Phoma betae and its Application in Synthetic Dyes Decolorization
1Botany and Microbiology Department, Faculty of Science, Cairo University, 12613, Giza, Egypt
2Bioprocess Development Department, Genetic Engineering and Biotechnology Research Institute (GEBRI), City for Scientific Research and Technology Application, New Borg El-Arab, 21934, Alexandria, Egypt
3Plant Protection and Biomolecular Diagnosis Department, Arid Lands Cultivation Research Institute, City for Scientific Research and Technology Application, New Borg El-Arab,21934, Alexandria, Egypt
Abstract
LACCASE (EC 1.10.3.2, p-diphenol: dioxygen oxidoreductases) is an extracellular enzyme found mainly in bacteria, fungi and plants. It has the ability to decolorize the hazardous synthetic dyes. Our study aimed to produce, purify, characterize and immobilize laccase from the fungus Phoma betae and its application in decolorization of synthetic dyes. Laccase was produced using both submerged and solid-state fermentations. Some inducers (ferulic acid, vertyl alcohol and CuSO4) for laccase production were used singly and in mixture. Significant increase in laccase production with all tested compounds was detected. Submerged fermentation induced laccase production than solid-state fermentation. Fast protein liquid chromatography (FPLC) used for laccase purification and through Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS/PAGE) the molecular mass of the purified laccase was 37 kDa. Optimum temperature for lacasse activity was determined at 30°C using guaiacol assay. Enzyme was more stable at -4ºC than at -20 and -80ºC. Maximum laccase activity was found at pH 8. Effect of metal ions and inhibitors on laccase activity was studied. Ni2+ and K1+ induced laccase activity, while Na1+, Ag1+, Hg2+, Zn2+, Cu2+, EDTA and SDS decreased laccase activity with different ratios. Laccase was immobilized in alginate beads. Both free and immobilized laccase were used to decolorize five synthetic dyes (Eriochrome Cyanine R, Merantine Brilliant Yellow 8G, Rhodamine 123, Solvent Orange 20 and Solvent Yellow 47) at concentration 0.2 g dye/L. Immobilized laccase decolorized dyes with higher yields than free one in all tested dyes. Phoma betae laccase is an efficient biocatalyst for synthetic dye decolorization.
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