Effect of salts and surfactants on the partitioning of Fusarium solani pisi cutinase in aqueous two-phase systems of thermoseparating ethylene oxide/propylene oxide random copolymer and hydroxypropyl starch.
An aqueous two-phase system composed by a thermoseparating random copolymer of ethylene oxide/propylene oxide 50/50 (%w/w), Breox, and hydroxypropyl starch-Reppal PES 100 was evaluated for the partitioning of Fusarium solani pisi recombinant cutinase. The effect of several additives on the partitioning of pure cutinase was evaluated. Micelles of sodium dodecanoate provided a ten-fold increase of the partitioning coefficient (K=9) and recovery yields of 60-75%. The phase diagrams of the systems composed of Breox, Reppal and sodium dodecanoate were determined and it was found that in systems with high surfactant concentrations, the binodal was moved to lower polymer concentrations, enabling a two-phase system with 6% (w/w) of each polymer.
Cunha
M T
MT
Centro de Engenharia Biológica e Química, Instituto Superior Técnico, Lisboa, Portugal.
Cabral
J M
JM
Tjerneld
F
F
Aires-Barros
M R
MR
D002021:Buffers; D002265:Carboxylic Ester Hydrolases / Q000302:isolation & purification*; D005670:Fusarium / Q000201:enzymology*; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D007850:Lauric Acids; D008823:Micelles; D011095:Polyethylenes; D011126:Polypropylenes; D011994:Recombinant Proteins / Q000302:isolation & purification; D012492:Salts; D013213:Starch; D013501:Surface-Active Agents
D002021:Buffers; D007850:Lauric Acids; D008823:Micelles; D011095:Polyethylenes; D011126:Polypropylenes; D011994:Recombinant Proteins; D012492:Salts; D013501:Surface-Active Agents; C030358:lauric acid; D013213:Starch; C078661:UCON 50-HB-5100; D002265:Carboxylic Ester Hydrolases; C019952:cutinase