Purification and Characterization of the Enzymes Lipoxygenase and Tyrosinase that Involved in the Quality of Desert Truffles (Terfezia claveryi)

Mohammed H. Mushrif; Abdullah E. Jasim; Luai F. Zghair

Purification and Characterization of the Enzymes Lipoxygenase and Tyrosinase that Involved in the Quality of Desert Truffles (Terfezia claveryi)

Mohammed H. Mushrif, Abdullah E. Jasim, Luai F. Zghair

Abstract

This contribution presents a review of the purification and characterization of two enzymes involved in the quality of desert truffles: lipoxygenase and tyrosinase. Both enzymes were extracted using phase partitioning in Triton X-114, a method which permits the elimination of most of the lipids and phenols and to obtain a discrete degree of purification. Tyrosinase from T. claveryi is a fully latent enzyme that was activated by the anionic surfactant SDS. The use of SDS also permitted the histochemical localization of the latent enzyme within the ascocarp. Tyrosinase was kinetically characterized using L-DOPA as
substrate. The effect of different inhibiting agents was also studied, tropolone being the most effective. The results obtained indicate that lipoxygenase from T. claveryi ascocarps is a soluble enzyme with a molecular weight of 66 kDa, which displays its maximum activity at pH 7.0 using linoleic acid as substrate. When using this fatty acid, the product of this reaction was the corresponding 13-hydroperoxide.

Keywords

purification, lipoxygenase tyrosinase, desert truffles

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