Purification And Characterization Of A Hyaluronidase From The Stingrays (Potamotrygon motoro) Venom. 

¹Magalhães, M. R., ²Ulhoa, C. J. and ¹Silva Jr., N. J.

1 Centro de Estudos e Pesquisas Biológicas, Univesidade Católica de Goiás; 2 Instituto de Ciências Biológicas, Universidade Federal de Goiás.

Freshwater stingray venom presents toxic fractions with hyaluronidase, acid phosphomonoesterase, 5´-nucleotidase, phospholipase activity and proteolytic activity on casein, gelatin and elastin. Hyaluronidases are enzymes that catalyze the cleavage of internal glycosidic bonds of certain mucopolysaccharides of animal connective tissues (e.g. hyaluronic acid). This enzyme are frequently referred as “spreading factor”, because hydrolysis of hyaluronic acid facilitates toxin diffusion into the tissues of the victim. In the present study, the hyaluronidase from the venom of Potamotrygonmotoro was purified and partialy characterized using chromatography with Sephacryl S-100 and SP-Sepharose. The enzyme was purified 376,4 times with an approximate molecular weight of 79 kDa and specific activity of 1,35 x 10⁸ NFU/mg. Maximum activity was demonstrated in pH 4,2 at 40ºC. The KM was estimated in 11,7 mg/ml and V_Maxin 33,69 mg of hyaluronic acid hydrolized per minute. The hyaluronidase of P. motoro was demonstrated to be heat instable, being stabilized by bovine albumin 1% and DTT, and inhibited by MnSO4. Since the venom of stingrays is just used for the defense, a more detailed investigation on the significance of a hyaluronidase so active will be necessary.

Correspondence to: martarm@terra.com.br