Purification, Characterization and in vitro Anthelmintic Activity of a Neutral Metalloprotease from Laccocephalum mylittae

Autor: Yongquan Li, Zhenxing Zhou, Yi-Qiong Zhang, Wen-Jun Guan, Qinqin Xu, Li-Hua Zhou
Rok vydání: 2010
Předmět:
Zdroj: Chinese Journal of Chemical Engineering. 18:122-128
ISSN: 1004-9541
DOI: 10.1016/s1004-9541(08)60332-8
Popis: A neutral metalloprotease was purified from the cultured mycelia of Laccocephalum mylittae , an effective medicinal fungus widely used in anthelmintic therapy. The protease was purified to homogeneity with 31.85-fold purification and a final yield of 21.76%. The subunit molecular weight of the protease is about 40000 estimated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The optimum reaction pH and temperature are 7.5 and 50°C, respectively. The protease activity is largely enhanced by Ca 2+ , but highly inhibited by tetrasodium ethylenediaminetetraacetate (EDTA), a metal-chelator, suggesting that the enzyme is a metalloprotease. The Michaelis-Menten constan K m and V max value for casein substrate are 6.09 mg·ml −1 and 21.32 μg·min −1 ·ml −1 , respectively. In vitro anthelmintic tests of the protease exhibit distinct lethal effects on the third stage larvae (L3) of Ascaris suum . Scanning electron microscopy and SDS-PAGE analysis indicates that the proteolysis of larvae proteins caused by this protease may relate to the anthelmintic activity of L. mylittae .
Databáze: OpenAIRE