Specificity of soybean lipoxygenase-1 in hydrated reverse micelles of sodiumbis(2-ethylhexyl)sulfosuccinate (aerosol OT) |
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| Authors: | Tatyana N Shkarina Hartmut Kühn Tankred Schewe |
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| Affiliation: | (1) Institute of Biochemistry of the Medical School (Charité), Humboldt University, Hessische Str. 3-4, D-O-1040 Berlin, Germany;(2) Present address: All-Union Vitamin Research Institute, Nauchny proezd 14a, 117820 Moscow, Russia |
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| Abstract: | Soybean lipoxygenase-1 (EC 1.13.11.33) was purified by fast protein liquid chromatography on a MONO Q column and studied with
respect to the conversion of linoleic and arachidonic acids in reverse micelles of sodiumbis(2-ethylhexyl)sulfosuccinate inn-octane. In this system the specific activities were lower by one order of magnitude than those in the corresponding aqueous
system. High-performance liquid chromatography analyses indicated the predominant formation of 13S-hydroperoxy-9Z,11E-octadecadienoic acid (13-HpODE) from linoleic acid and of 15S-hydroperoxy-5Z,8Z,11Z,13E-eicosatetraenoic acid (15) from arachidonic acid both in the aqueous system and in the reverse micelles. After sedimentation
of the hydrated reverse micelles by ultracentrifugation, both linoleic acid and 13-HpODE were found to be enriched in the
micelles with only small amounts of these compounds present inn-octane. It is proposed that substrates and products of the lipoxygenase reaction are located mainly in the surfactant shell
of the hydrated reverse micelles and reach the micelle-entrapped enzyme by diffusion into the aqueous interior space. |
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