Optimization of Recombinant Pseudomonas putida Proline Dehydrogenase Purification in Aqueous Two-Phase systems Using Response Surface Methodology

Nasrin Masoudzadeh, Eskander Omidinia
Protein Expression and Purification 2014 April 29
An experimental design was employed to optimize the purification conditions of a recombinant proline dehydrogenase (ProDH) from Pseudomonas putida which over-expressed in E. coli as insoluble inclusion bodies. Integration of solubilization, reconstitution and extraction in aqueous two-phase systems (ATPS) were employed as a method for recovery of recombinant ProDH. To build the mathematical model and reduce the processing steps of experiments trials necessary to evaluate optimal conditions, response surface methodology (RSM) with a central composite rotatable design (CCRD) was defined. The adequacies of the calculated models for the responses were evaluated by means of variance analysis. The optimal extraction conditions were found using the 14.33% PEG 6000, 11.79% sodium carbonate at pH 7.48. The agreement between predicted and experimental results verifies the validity of the response model.

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