Extraction of epoxide hydrolase from Glycine max using microwave-assisted three phase partitioning with dimethyl carbonate as green solvent

Abstract

Epoxide hydrolase from Glycine max was partially purified and recovered in a single step using three phase partitioning (TPP). It was found to be exclusively partitioned in the interfacial precipitate phase. TPP was explored for the extraction of epoxide hydrolase using dimethyl carbonate as an organic phase and ammonium sulfate as a salt phase. It was then combined with microwave irradiation for process intensification, and the effect of microwave irradiation on the extent of extraction recovery was studied. The % percent recovery and purification fold achieved in TPP were 121.2 ± 1.6 % and 2.14 ± 0.03, respectively, under the optimized conditions of 40 % ammonium sulfate precipitation, 1:1.5 (v/v) crude extract to solvent ratio at 45 °C. These optimized conditions were further operated under microwave irradiation reducing the contact time from 1 h to 8 min with 20 W microwave power and 50 % duty cycle achieving recovery of 142.6 ± 1.9 % and purification fold of 2.38 ± 0.02. The percent recovery was enhanced up to 1.17 times with the use of microwave-assisted TPP (MTPP). The combined microwave-assisted TPP was found to be a green and effective technique for the recovery and partial purification of epoxide hydrolase from Glycine max.