Adsorptive immobilization of intestinal brush border membrane on Triton X-100-substituted Sepharose 4B

Triton X-100-substituted Sepharose 4B (Sepharose-TX) was used for adsorptive immobilization of intestinal brush border membrane using lactose-phlorizin hydrolase as a representative membrane enzyme. Limited heating of membrane preparations was found to enhance binding. This enhancement is concluded to be owing to a greater availability of the hydrophobic sites, as also confirmed by the 1-anilino-8-naphthalene sulfonate fluorescence studies, for interaction with Triton X-100 moieties on the support. The immobilized preparations obtained by this procedure were found useful in hydrolysis of lactose, involving lactose-phlorizin hydrolase, in continuous operations. It is suggested that the approach may be of general utility for immobilization of biologic membranes by interaction of their extramembrane structures using supports with appropriate hydrophobic groups.