The influence of lipid matrixes with different charges on the protein folding behavior has been investigated by using the Langmuir-Blodgett (LB) technique. To understand the conformation change under different forces, Glucose oxidase (GOD) from Aspergilius Niger was used as a protein model. Eight glycolipids(1,2-o-dialkyl-3-o-beta-D-glycosylglycerols and alkyl 2-amino-2-deoxy-beta-D-glucopyranoside) were used as the matrixes for this investigation. It was observed that the GOD can penetrate into neutral glycolipid monolayer and change its conformation in favor of the alpha-helix formation. Moreover, GOD strongly adsorbed to the positive charged glycolipid monolayer and change its conformation in favor of the beta-sheet formation. Enzymatic activity measurements showed that the more the alpha-helix conformation content is in the GOD, the higher activity the GOD will be. This fact suggested a new way to mediate the conformation of protein in organized molecular assemblies, and provided a new thinking for the preparation of biomimetic film and biosensor.