Studies on the Enzymes of Sarcocystis suicanis: Purification and Characterization of an Acid Phosphatase

Abstract

Percoll density gradient centrifugation was used for isolating large quantities of bradyzoites of Sarcocystis suicanis, which were used for enzymatic analysis. Crude extracts of bradyzoites contained activities suggestive of several acid hydrolases. Levels of acid and alkaline phosphatase were higher than those of beta-N-acetylhexosaminidase and beta-galactosidase. Acid phosphatase was purified 156-fold with an overall recovery of 54% using DEAE-Sepharose 4B and Sephadex G-200 chromatography. The partially purified enzyme was not a glycoprotein and had a molecular weight of approximately 170,000. The enzyme was markedly inhibited by Cu++, Hg++, and iodoacetamide, suggesting the presence of a sulfhydryl group. Sodium tartrate caused strong inhibition of the enzyme. The acid phosphatase of S. suicanis appears to be a unique enzyme that cannot be classified under high or low molecular weight acid phosphatases of widely diverse origin.

Partial purification and biochemical properties of acid and alkaline phosphatases from Myxococcus coralloides D

“…The acid phosphatase produced by M. coralloides D cannot be placed in either of the above categories, since it resembles the high molecular weight group, being insensitive to tartrate and sulphydryl agents but it also has a broad substrate specificity. This situation has also been described in the acid phosphatase from Sarcocystis surcanis (Farooqui et al 1987).…”

Isolation, Culture and Cryopreservation of Sarcocystis species

“…The objective is to release bradyzoites from sarcocysts that were broken during grinding or homogenization, and this method is best suited for heavily infected tissues. Lightly infected tissues may have to be digested longer in digestion fluid, and in that case, the digestion fluid should be isotonic (Farooqui et al, 1987).…”

“…The number of sarcocysts in different tissues may vary among animals; heart, and tongue are the most heavily infected organs (Dubey et al, 2016). The following method is useful for obtaining bradyzoites in sufficient quantity for biological and molecular characterization (Farooqui, Adams, Hanson, & Prestwood, 1987;Lunde & Fayer, 1977). 2.…”

Macrophage responses to Toxoplasma antigens in vitro: a possible role in inflammatory lesions in toxoplasmosis.

“…Acid phosphatase activity was measured by a modified method of Farooqui and others. 20 Two hundred microliters of 15 mM p-nitrophenyl phosphate dissolved in acetate buffer were added to 150 l of acetate buffer (pH 4.5) in a reaction vessel. One hundred microliters of supernatant or cellular extract adjusted to contain 30 g/ml of protein were added to the mixture and incubated at 37ЊC for 15 min.…”