Skip to main content
. Author manuscript; available in PMC: 2007 Oct 1.
Published in final edited form as: Protein Expr Purif. 2006 Dec 1;52(2):334–339. doi: 10.1016/j.pep.2006.11.009

Table 1.

Purification of recombinant WNV NS2B-NS3pro and NS2B-NS3pro K48A from E. coli cell extract1

Total protein, mg Total activity, U2 Specific activity, U/mg Purification, -fold Yield, %
WT K48A WT K48A WT K48A WT K48A WT K48A
Crude extract 248 234 26.2 25,7 0.106 0.11 1 1 100 100
Chromatography 47.4 43.7 18.1 18.9 0.38 0.43 3.6 3.9 69.1 73.5
Re-chromatography 11.2 13.1 10,7 12 0.9 0.9 8.5 8.2 40.8 46.7
5 kDa-cut off concentration 10.5 12 10.2 11.5 1 1 9.4 9.1 38.9 44.7
1

E.coli cells (5 g wet weight) were used for purification. NS2B-NS3pro, WT; NS2B-NS3pro K48A, K48A.

2

One unit (U) of enzymatic activity corresponds to the liberation of 1 μmol of 7-amino-4-methylcoumarin (AMC) per min/mg from the Pyr-RTKR-AMC substrate.