Development and Application of Aqueous Two-Phase Partition for the Recovery and Separation of Recombinant Phenylalanine Dehydrogenase

作者：Mohammadi Shahbaz Hamid; Omidinia Eskander^*

来源：Iranian Journal of Chemistry and Chemical Engineering (International English Edition), 2008, 27(2): 119-127.

• Aqueous two-phase systems (ATPS)
• Ammonium sulfate
• Phenylalanine dehydrogenase (PheDH)
• Polyethylene glycol (PEG)
• Separation

Aqueous two-phase systems (ATPS) have emerged as a powerful extraction method for the downstream processing of bio-molecules. The aim of this work was to investigate the possibility of utilizing ATPS for the separation of recombinanat Bacillus sphaericus phenylalanine dehydrogenase (PheDH). Polyehtylene glycol (PEG) and ammonium sulfate systems were selected for our experiment. The effect of different elements such as: type and concentration of PEG, concentration of (NH(4))(2)SO(4) pH, phase volume ratio (VR) and tie-line length (TLL) on the extraction behavior and selective separation was also studied. Desirable conditions for differential partitioning was obtained in 8.5 % (w/w) PEG-6000, 17.5 % (w/w) (NH(4))(2)SO(4) and V(B) 0.25 at pH 8.0. PheDH was mainly concentrated into the upper PEG-rich phase in all tested systems. The partition coeffficient (K), recovery (R %), yield (Y %). TLL and selectivity were found to be 58.7, 135 %, 94.42 %, 39.89 % (w/w) and 2174, respectively. From the experimental results, it was revealed that the PEG molecular weight, (NH(4))(2)SO(4) concentration. TLL and pH of system had strong impacts on partition features. The extraction efficiency was increased with elevation of pH and TLL values. In this paper, we described the partitioning behavior in PEG/(NH(4))(2)SO(4) ATPS in order to evaluate the applicability of ATPS for partitioning and recovery of PheDH.

• 出版日期2008