خالص‌سازی آنزیم لاکاز قارچ Trametes و تعیین خصوصیات فیزیکوشیمیایی لاکاز نوترکیب: یک مطالعه آزمایشگاهی

Background and Objectives: Laccase is the most abundant member of protein family that catalyzes the oxidation of substituted phenols. Laccases are used as biocatalysts for decolorization and bleaching in dye industries, detoxification in environment, and juice clarification in food industries. The present study aimed at producing recombinant laccase, purifying with high yield and fold, and characterizing biochemical and kinetic properties of the purified laccase. Materials and Methods: In this laboratory study, recombinant Trametes laccase, which was successfully expressed in yeast host in extracellular form, was purified through ultrafiltration and gel chromatography methods. The purification process yield and fold was determined and after molecular weight determination, physiochemical properties of recombinant laccase were studied and its thermostability was determined. All tests were conducted with at least 3 times repetition and mean and standard deviation calculation.   Results: Purification strategy showed that laccase was successfully purified with 5.67 fold and 49.7% yield and the molecular weight of recombinant laccase was determined 65KDa. Optimum pH and temperature of the recombinant laccase were determined 4.8 and 60°C, respectively. Its kinetic properties were also determined. The thermostability assessment showed that purified laccase is highly thermostable. Conclusion: The results indicated that gel chromatography purification method can be an ideal to achieve high yield and fold purification of recombinant laccase with keeping enzyme properties and high thermostability, so that the recombinant purified laccase with high yield and fold can be used in drug, textile, and food industries, and environment and its detoxification.   Key words: Purification, Laccase enzyme, Trametes fungal, Physicochemical properties   Funding: This study was funded by Islamic Azad University, Central Tehran Branch. Conflict of interest: None declared.   How to cite this article: Hesampour A, Mohandesi N. Purification of Fungal Laccase from Trametes and Characterization of Recombinant Laccase Physicochemical Properties. J Rafsanjan Univ Med Sci 2018; 17 (6): 511-22. [Farsi]