Ellipsometric study of molecular orientations of Thermomyces lanuginosus lipase at the air–water interface by simultaneous determination of refractive index and thickness

• In-situ detection of lipase orientations at the liquid/fluid interface.
• Monolayer adsorption kinetics monitored with subnanometer resolution.
• pH value and concentration determines orientation of the macromolecules.
• Compression beyond a threshold induces reorientation of the molecules.

Ellipsometric studies of very thin organic films suffer from the low refractive index contrast between layer and bulk substrate. We demonstrate that null ellipsometry can not only provide detailed information about the adsorption kinetics and surface excess values, but in addition on layer thicknesses with submonolayer resolution of a lipase from Thermomyces lanuginosus at the air–water interface. While measuring very close to the Brewster angle, refractive indices and layer-thicknesses can both be determined with a precision that is sufficiently high to make conclusions on the density and orientation of the molecules at the interface. The orientation was found to be concentration- and pH value-dependent. At the isoelectric point, the lipase was almost vertically oriented with respect to the surface, while for pure distilled water and low lipase concentration a rather horizontal alignment was found. Further experiments, varying the size of the interfacial area in a Langmuir trough, confirm the different layer structures.

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