Purification And Characterization Of G . M. Morsitans Midgut Trypanolysin

Abstract

In the present study, a trypanolysin induced by component of blood meal in the midgut of tsetse G. m. morsitans was isolated and purified in three steps. The first step was achieved by separation of the midgut homogenate on a conventional anion-exchange chromatography column. The highest trypanolysin activity was recovered in the bound fractions (95%, 05 M NaCl). In the second step, isolation was achieved on a MonoQ anion-exchange column by elution at 70 -80% 1 M NaCl. The third step of solation-purification was achieved by using epoxy-activated Sepharose 6-B-affinity chromatography column.In this case, the trypanolysin was eluted using 20 mM Tris-HCl.The purified native G. m. morsitans trypanolysin was of Mw≈669 kDa, while ≈ 14 kDa trypanolysin was shown in denaturing trypanolysin SDS -PAGE gel for the same tsetse species. It was noticed that the purified trypanolysin was lipidated and also found to be glycosylated.