Extraction of Papain by Ionic Liquid/Aqueous Two-phase System and Optimization of Process Conditions
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Abstract:
Papain was extracted by a [C4mim] Br/K2HPO4 ionic liquid/aqueous two-phase system. First, the influence of reagents on the activity of papain was investigated. Second, the effects of alkyl chain length and concentration of the ionic liquid, dosage of papain, pH, and temperature on the partitioning behavior of papain were discussed concretely, and the extraction conditions were optimized using the central composite design (CCD). The results showed that ionic liquids had little impact on the activity of papain and could promote the activity at low concentrations, while the inorganic salt had a larger influence on the activity of papain. There was a negative?influence on the extraction of papain in the ionic liquid/aqueous two-phase system at high temperature. The influence of the various parameters was in the following order: concentration of K2HPO4 > concentration of [C4mim]Br>dosage of papain> pH. The optimized extraction conditions were as follows: [C4mim] Br content, 0.30 g/mL; K2HPO4 content, 0.30 g/mL; pH, 6.0; dosage of papain, 3.0 mg/mL; reaction temperature, 30.0 ℃. Under the optimum conditions, the activity recovery of the enzyme and purification factor reached 91.20% and 1.73, respectively. The result provided the experimental basis for further research and large-scale production.
