A hyaluronidase from Potamotrygon motoro (freshwater stingrays) venom: isolation and characterization.
Freshwater stingrays (Potamotrygon motoro) are known to cause human accidents through a sting located in its tail. In the State of Goiás, this accident happens especially during the fishing season of the Araguaia River. The P. motoro venom extracted from the sting presented hyaluronidase activity. The enzyme was purified by gel filtration on Sephacryl S-100 and ion-exchange chromatography on SP-Sepharose. A typical procedure provided 376.4-fold purification with a 2.94% yield. The molecular weight of the purified enzyme was 79 kDa as estimated by gel filtration on Sephacryl S-100. The K(m) and V(max) values for hyaluronidase, using hyaluronic acid as substrate, were 4.91 microg/ml and 2.02 U/min, respectively. The pH optimum for the enzyme was pH 4.2 and maximum activity was obtained at 40 degrees C. The hyaluronidase from P. motoro was shown to be heat instable, being stabilized by bovine albumin and DTT, and inhibited by Fe(2+), Mn(2+), Cu(2+) and heparin.
Magalhães
Marta R
MR
Centro de Estudos e Pesquisas Biológicas, Departamento de Biologia, Universidade Católica de Goiás, 74.605-010, Goiânia, GO, Brazil.
da Silva
Nelson Jorge
NJ
Ulhoa
Cirano J
CJ
D000818:Animals; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003300:Copper / Q000494:pharmacology; D004229:Dithiothreitol / Q000494:pharmacology; D018940:Elasmobranchii; D004591:Electrophoresis, Polyacrylamide Gel; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005618:Fresh Water; D006493:Heparin / Q000494:pharmacology; D006358:Hot Temperature; D006820:Hyaluronic Acid / Q000378:metabolism; D006821:Hyaluronoglucosaminidase / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007501:Iron / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D013379:Substrate Specificity; D014688:Venoms / Q000201:enzymology*
D004791:Enzyme Inhibitors; D014688:Venoms; D012710:Serum Albumin, Bovine; D008345:Manganese; D003300:Copper; D006820:Hyaluronic Acid; D006493:Heparin; D007501:Iron; D006821:Hyaluronoglucosaminidase; D004229:Dithiothreitol