title: Development of the simple and sensitive method for lipoxygenase assay in AOT/isooctane reversed micelles.
abstract: In this study, we investigated the possibility of reversed micelles, widely used as an enzyme reactor for lipases, for the determination of lipoxygenase activity. Although it is rapid and simple, reversed micelles have some limitations, such as interference by UV-absorbing materials and surfactant. Lipoxygenase activity in the reversed micelles was determined by reading the absorbance of the lipid hydroperoxidation product (conjugated diene) at 234 nm. Among surfactants and organic media, AOT and isooctane were most effective for the dioxygenation of linoleic acid in reversed micelles. The strong absorbance of AOT in the UV region is a major obstacle for the direct application of the AOT/isooctane reversed micelles to lipoxygenase activity determination. To prevent interference by AOT, we added an AOT removal step in the procedure for lipoxygenase activity determination in reversed micelles. The lipoxygenase activity was dependent on water content, and maximum activity was obtained at an R-value of 10.
authors: Department of Agricultural Biotechnology, Seoul National University, Seoul 151-742, Republic of Korea.
authors: Kyung Min
authors: KM
authors: Park
authors: Yu Na
authors: YN
authors: Kim
authors: Seung Jun
authors: SJ
authors: Choi
authors: Pahn-Shick
authors: PS
authors: Chang
mesh_terms: D057075:Enzyme Assays / Q000379:methods*; D007700:Kinetics; D008084:Lipoxygenase / Q000032:analysis*; D008823:Micelles; D009795:Octanes / Q000737:chemistry; D012680:Sensitivity and Specificity; D013386:Succinates / Q000737:chemistry; D013501:Surface-Active Agents / Q000737:chemistry
chemical_list: C512554:1,4-bis(2-ethylhexyl) sodium sulfosuccinate; D008823:Micelles; D009795:Octanes; D013386:Succinates; D013501:Surface-Active Agents; D008084:Lipoxygenase; C045798:2,2,4-trimethylpentane