Dynamic light scattering of cutinase in AOT reverse micelles.
The fungal lipolytic enzyme cutinase, incorporated into sodium bis-(2ethylhexyl) sulfosuccinate reversed micelles has been investigated using dynamic light scattering. The reversed micelles form spontaneously when water is added to a solution of sodium bis-(2ethylhexyl) sulfosuccinate in isooctane. When an enzyme is previously dissolved in the water before its addition to the organic phase, the enzyme will be incorporated into the micelles. Enzyme encapsulation in reversed micelles can be advantageous namely to the conversion of water insoluble substrates and to carry out synthesis reactions. However protein unfolding occurs in several systems as for cutinase in sodium bis-(2ethylhexyl) sulfosuccinate reversed micelles. Dynamic light scattering measurements of sodium bis-(2ethylhexyl) sulfosuccinate reversed micelles with and without cutinase were taken at different water to surfactant ratios. The results indicate that cutinase was attached to the micellar wall and that might cause cutinase unfolding. The interactions between cutinase and the bis-(2ethylhexyl) sulfosuccinate interface are probably the driving force for cutinase unfolding at room temperature. Twenty-four hours after encapsulation, when cutinase is unfolded, a bimodal distribution was clearly observed. The radii of reversed micelles with unfolded cutinase were determined and found to be considerable larger than the radii of the empty reversed micelles. The majority of the reversed micelles were empty (90-96% of mass) and the remainder (4-10%) containing unfolded cutinase were larger by 26-89 A.
Unidade de Ciências e Tecnologias Agrárias, Universidade do Algarve, Campus de Gambelas, 8000, Faro, Portugal. emelo@ualg.pt
E P
EP
Melo
P
P
Fojan
J M
JM
Cabral
S B
SB
Petersen
D002265:Carboxylic Ester Hydrolases / Q000737:chemistry*; D004143:Dioctyl Sulfosuccinic Acid; D008027:Light; D008823:Micelles; D012542:Scattering, Radiation
D008823:Micelles; D004143:Dioctyl Sulfosuccinic Acid; D002265:Carboxylic Ester Hydrolases; C019952:cutinase