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Functional characterization of X-prolyl aminopeptidase from Toxoplasma gondii.

Parasitology. 2016-10; 
YangMingfa,ZhengJun,JiaHonglin,SongMin
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Biochemicals … Purification of the rTgAPP was performed with Glutathione Resin (GenScript, USA) according to the manufacturer's instructions. The rTgAPP fused with GST was eluted with 20 mM reduced glutathione (GE Health care, Piscataway, USA) … Get A Quote

摘要

In the present study, a recombinant aminopeptidase P (rTgAPP) from Toxoplasma gondii was expressed in Escherichia coli to evaluate its enzyme parameters. The rTgAPP showed strong activity against a synthetic substrate for aminopeptidase P at pH 8·0 with a K m value of 0·255 µ m and a k cat value of 35·6 s-1. The overall catalytic efficiency (k cat/K m) of the rTgAPP was 139·6 × 105 M-1 s-1. The activity of rTgAPP was enhanced by the addition of divalent cations and inhibited by bestatin. Deletion of TgAPP gene in the parasite through a CRISPR/Cas9 system resulted in inhibition of growth indicating the importance of TgAPP. Thus our findings reveal that TgAPP is an active enzyme in T. gondii and provide a... More

关键词

Toxoplasma gondii,Toxoplasmosis,X-prolyl aminopeptidase,enzymatic activity,gr