Removing Unwanted Protein Tags Recombinant Bovine Enterokinase REK
YaxinBio Enterokinase is a kind of highly purified recombinant
bovine enterokinase. The enzyme has been extensively purified and
there are no traces of other contaminating proteases. Enterokinase
specifically hydrolyzes peptide bond at the carboxyl side of lysine
residue preceded by four aspartic acids: Asp-Asp-Asp-Asp-Lys
(DDDDK). So, Enterokinase can remove N-terminal fusion protein or
tags to get aim protein with native amino acids sequence.
Source: bovine enterokinase, expressed in E. Coli
Common components influence the action of enterokinase
＞200mM imidazole or ＞200mM NaCl or ＞5%glycerin, the reaction may be
effected.The following suggestions are given:
1) To receive the optimum result, please dialyze the sample to 25
mMTris-HCl, pH 8.0.
2) If the dialysis is inconvenient, please dilute the sample to
＜100mM imidazole, ＜50mMNaCl, ＜5% glycerin, and the proportion of
fusion protein and EK may not be changed (1U:0.5mg fusion protein).
3) If there are one or more components in samples, and cannot be
removed, suggest to increase the content of EK in reaction system
or extend the reaction time.
Main Features Advantages
One unit is defined as the amount of enzyme needed to cleave 0.5mg
of fusion protein in 12 to16 hours to get 95% completion at 25°C in
25mMTris-HCl, pH 8.0. Substrate: a special fusion protein.
-20°C or below.
Keep cool with blue ice during shipping. Remained stable at 25°C
for one week without activity lost. No activity lost after 5 cycles
Enterokinase is a member of the S1 peptidase family. In vivo, it is responsble for the proteolytic activation of trypsin from
trypsinogen. Enterokinase is used for site specific cleavage of
recombinant fusion proteins containing an accessible enterokinase
recognition site for removal of affinity tags.
Enterokinase from bovine intestine has been used in a study to
assess duodenase as a potential activator of cascade digestive
proteases. Enterokinase from bovine intestine has also been used in
a study to investigate an inhibitor of enteropeptidases and trypsin
from the bovine duodenum.
The enzyme has been used to compare the specific activity with that
of purified, recombinant bovine enterokinase (light chain)
overexpressed inEscherichia coli.