Process Biochemistry
Process Biochemistry
4 years ago

Application of microwave assisted three phase partitioning method for purification of laccase from Trametes hirsute

Application of microwave assisted three phase partitioning method for purification of laccase from Trametes hirsute
A novel application of microwave assisted three phase partitioning (MTPP) was studied for the first time in partial purification of laccase from Trametes hirsuta MTCC-1171. Conventional three-phase partitioning (TPP) was optimized and used as a reference method, which yielded 53.9±0.9% of enzyme with 2.4±0.07-fold purity with optimized experimental conditions such as ammonium sulfate concentration (40% w/v), crude to tert-butanol ratio (1:1v/v), pH 4.5, and temperature 30°C. A Box-Behnken design (BBD) of response surface methodology (RSM) was exercised to optimize MTPP. With RSM optimized experimental conditions of MTPP such as microwave irradiation time (7min), duty cycle (20%), and power (20W), the maximum yield obtained was 59.8±1.4% with purity of 13.9±0.5-folds which coincides well with the predicted values. Laccase purity and yield were enhanced within a considerably less processing time (7min in MTPP as against 60min in conventional TPP). However, various experimental parameters showed adverse effects on laccase partitioning when higher values of power, exposure time and duty cycle were applied. SDS-PAGE analysis confirmed the partitioning of partially purified laccase from crude broth with a molecular weight in the range of 67–72kDa.

Publisher URL: www.sciencedirect.com/science

DOI: S1359511317313193

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