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LOXPsa1, the first recombinant lipoxygenase from a basidiomycete fungus

Plagemann, Ina, Zelena, Katerina, Arendt, Philipp, Ringel, Peter D., Krings, Ulrich, Berger, Ralf G.
Journal of Molecular Catalysis. B, Enzymatic 2013 v.87 pp. 99-104
linoleic acid, Pleurotus sapidus, hydrocarbons, linoleate 13S-lipoxygenase, odors, high performance liquid chromatography, amino acid sequences, pH, temperature, enzyme activity, catalytic activity, hydroperoxides, grapefruits, fungi
A dioxygenase from the edible basidiomycete Pleurotus sapidus, originally researched because of its distinct ability to convert the sequiterpene (+)-valencene to the valuable grapefruit aroma (+)-nootkatone, was identified as a potent lipoxygenase (LOXPsa1). Kinetic parameters, pH and temperature optima of the pure recombinant enzyme were determined using linoleic acid as the substrate. Km, vmax, and kcat were 40.3μM, 130.3U mg−1, and 157s−1, respectively. The maximal enzymatic activity was found at pH 7.0 and 35°C. Showing high specificity toward free linoleic acid, the enzyme was classified as lipoxygenase type 1. Conversion of linoleic acid yielded mainly (S)-13-hydroperoxy-9Z,11E-octadecadienoic acid (94% ee), as was confirmed by chiral HPLC analysis of the hydroperoxides. The amino acid sequence showed homology to lipoxygenases catalyzing S stereospecific oxygenation, and thus the enzyme was characterized as a 13S-lipoxygenase. This is the first lipoxygenase described to accept terpene hydrocarbons as substrates.