Tev Protease Application: Pharmaceutical Industry

Recombinant TEV Protease is a highly purified preparation of the catalytic domain of the small nuclear inclusion a (NIa) protein encoded by the etch virus (TEV). The biological function of the protease is to proteolyse the viral polyprotein into single proteins during etch virus biogenesis. Its sequence specificity is far more stringent than that of factor Xa, thrombin. TEV protease is a very useful tool enzyme for the downstream fusion proteins cleavage which recognizes a heptapeptide consensus sequence of the general form E-Xaa-Xaa-Y -Xaa-Q-P1AcA A (G/S), with cleavage occurring between Q and P1A A . The most commonly used sequence is ENLYFQG. Many different small amino acids are tolerated in the P1A A position, allowing target genes to be released from N-terminal fusions with either a native N-terminus or with only a single amino acid substitution. It has optimally engineered and purified for enhanced stability and high specific activity over a broad temperature range. The enzyme is produced in recombinant E.coli and purified to high homogeneity with specificity of 10,000U/mg. The optimal temperature for cleavage is 30AcA A , however, the enzyme can be used at temperatures as low as 4AcA A .
Purity: More than 95% by SDS-PAGE analysis.
Activity: One unit is defined as the amount of enzyme that will cleave 95% of 5A A g fusion protein in 12~16 hours at 4AcA A in 1x reaction buffer.
Recommended Enzyme Disgestion Conditions
Fusion protein0.1mg
Recombinant TEV protease20U
10xbuffer10
Temperature: 4
Total volume:10
Time A hour: 12~16
10x Reaction Buffer
500 mM Tris-HCl, pH 8.0
5 mM EDTA
1% Tween-20 (v/v)
10 mM DTT
Stability: Stable at -20AcA A for up to two years since reported data.
Reference
Kapust RB, TACr J, Copeland TD, Waugh DS. The P1' specificity of etch virus protease, Biochem Biophys Res Commun. 2002 Jun 28;294(5):949-55.