Enterokinase；Enteropeptidase CAS:9014-74-8

Product Description: enterokinase (light chain), CAS No.: 9014-74-8, is a highly specific protease that recognizes ASP ASP ASP ASP Lys sequence and hydrolyzes peptides at the C-terminal of Lys. It can convert trypsinogen into trypsin, or cut the fusion protein with this recognition sequence. Because of its high specificity and high hydrolysis efficiency, enterokinase is widely used in the development of genetic engineering products. One of its applications is as a tool protease for the specific cleavage of recombinant fusion proteins, especially in the bioengineering pharmaceutical industry and the research of genetic engineering, biochemistry and molecular biology.

This product is a bovine enterokinase with high purity and activity secreted and expressed by recombinant yeast. It has a wide range of application (4-45oc, pH 4.5-9.5), and still has partial activity in the presence of various descaling and denaturant.

Grade：Recombinant
Definition of enterokinase activity: the amount of enzyme required to degrade 95% of 0.5 mg of thioredoxin-np-27 to np-27 within 16 hours at 37oC is defined as 1 active unit

Properties: sterilized liquid, recombinant

Main function: for biochemical research. Enterokinase is a serine protease, which can efficiently and specifically hydrolyze the fusion protein expressed by engineering bacteria (the recognition order is - (ASP) 4lys ↓ --) and release the target protein. Enterokinase is widely used in the development of genetic engineering products because of its high specificity and high hydrolysis efficiency; It can be used as a protease to specifically identify and cleave the substrate containing enterokinase cleavage sites. One of its applications is as a tool protease for the specific cleavage of recombinant fusion proteins, especially for bioengineering pharmaceutical industry, genetic engineering, biochemistry, molecular biology and so on. However, the source of natural enterokinase is limited, and enterokinase extracted from animal tissues is contaminated with other proteases, which brings difficulties to practical application. This requires the production of high-purity enterokinase by genetic engineering. The activity of high-purity enterokinase produced by genetic engineering method is similar to that of natural enzyme, but the cleavage rate of enterokinase is faster.

Storage environment: - 20 ℃

There are many kinds of enterokinase chemical reagents, and the classification and grading standards of chemical reagents are different all over the world.

IUPAC classifies chemical reference materials as:

Class A: atomic weight standard.

Grade B: the reference material closest to Grade A.

Grade C: standard reagent with content of 100 + - 0.02%

Grade D: standard reagent with content of 100 + - 0.05%

Grade E: reagent with purity obtained by comparative determination with grade C or D as standard

Chemical reagents can be divided into standard reagents, general reagents, biochemical reagents, etc. In China, it is customary to refer to grade C and D reagents equivalent to IUPAC as standard reagents.

 Common factors affecting recombinant enterokinase activity:

>2M urea，>250mM NaCl，>20mM b-ME，>0.1% SDS，>50mM imidazole。
If the sample solution contains one or more of the above components, in order to obtain ideal enzyme digestion results, please dialysis the sample to 1 × In the reaction buffer, and then the enzyme digestion experiment was carried out.

Enzymatic digestion of recombinant bovine enterokinase:

Each reaction contains 15 μ G thioredoxin-np-27 and different amounts of enterokinase. After 16 hours of reaction at 37 ° C, the enzyme digestion effect was detected with SDS-PAGE gel at 15 ° C. The amount of enzyme contained in each reaction is:

Lane A: No rEK Lane                 B: 0.0008 units
Lane C: 0.0015 units Lane        D: 0.003 units
Lane E: 0.008 units Lane          F: 0.015 units Lane    G: 0.03 units

Recombinant bovine enterokinase