Tobacco Etch Virus (TEV) protease has widely used to cleave recombinant fusion protein. TEV protease is a 49 kDa nuclear inclusion A protein containing 27 kDa of catalytic domain. This catalytic domain of TEV protease is commercially purchased but its usage has been limited due to its auto-inactivation and cost. In here, we have isolated newly generated 6xhis-tagged S219V mutant catalytic domain of TEV protease in the background of T17S, N68D and I77V mutation engineered to improve soluble protein yield, and then measured its activity using 5-FAM fluorescence as a substrate. We found that V<sub>max</sub> for the isolated S219V mutant TEV protease was faster than that for commercially purchased one. Furthermore, new TEV protease did clearly cut off recombinant protein containing its cleavage site without self-inactivation. These results suggest that the mutation of S219 to Valine could improve TEV protease activity.

S219V mutation on tobacco etch virus (TEV) protease improved its activity

2019 Fall National Meeting

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