Partition features and renaturation enhancement of Chymosin in aqueous two-phase systems.

GEORGINA REH; DARIO, SPELZINI; GISELA ,TUBIO; GUILLERMO PICÓ; BEATRIZ FARRUGGIA

Journal of Chromatography B

ELSEVIER SCIENCE BV

Lugar: Amsterdam; Año: 2007 vol. 860 p. 98 - 105

1570-0232

Aqueous two phase systems of polyethyleneglycol-phosphate and polyethylen-polypropilen oxide maltodextrin systems were used in order to study the partition features of recombinant chymosin from inclusion bodies. Theses systems with 8M Urea were used for the solubilization and unfolding of inclusion bodies containing recombinant chymosin and the oxidative renaturation of this protein. Recombinant chymosin showed to be partitioned in favour of the top phase in all studied systems with a partition coefficient between 4 to 6. The solubilization-refolding process in urea of inclusion bodies was made directly in a poly ethylene-poly propylene oxides random copolymer-maltodextrin system and in the polyethyleneglycol-phosphate systems. The recovery of the chymosin biological activity was 32% in the first system while in the polyethyleneglycol-phosphate was (50-59 %). The results indicate that the liquid-liquid extraction would be an adequate tool able to isolate and concentrate chymosin from inclusion bodies with a yield of biological activity higher than the standard method.