GLK E.Coli, Active
Escherichia Coli.
Glucokinase, ECK2384, JW2385.
Greater than 95.0% as determined by SDS-PAGE.
Description
GLK E.Coli Recombinant produced in E.Coli is a single, non-glycosylated polypeptide chain containing 344 amino acids (1-321) and having a molecular mass of 37.1kDa.
GLK is fused to a 23 amino acid His-Tag at N-terminus and purified by proprietary chromatographic techniques.
Product Specs
Glucokinase, ECK2384, JW2385.
Escherichia Coli.
MGSSHHHHHH SSGLVPRGSH MGSMTKYALV GDVGGTNARL ALCDIASGEI SQAKTYSGLD YPSLEAVIRV YLEEHKVEVK DGCIAIACPI TGDWVAMTNH TWAFSIAEMK KNLGFSHLEI INDFTAVSMA IPMLKKEHLI QFGGAEPVEG KPIAVYGAGT GLGVAHLVHV DKRWVSLPGE GGHVDFAPNS EEEAIILEIL RAEIGHVSAE RVLSGPGLVN LYRAIVKADN RLPENLKPKD ITERALADSC TDCRRALSLF CVIMGRFGGN LALNLGTFGG VFIAGGIVPR FLEFFKASGF RAAFEDKGRF KEYVHDIPVY LIVHDNPGLL GSGAHLRQTL GHIL
Product Science Overview
Recombinant DNA technology involves inserting the gene encoding glucokinase into E. coli, which then expresses the enzyme. This method allows for the production of large quantities of the enzyme with high purity and activity. The recombinant glucokinase produced in E. coli is typically a single, non-glycosylated polypeptide chain containing 344 amino acids, with a molecular mass of approximately 37.1 kDa .
The enzyme is often tagged with a His-tag at the N-terminus to facilitate purification using affinity chromatography techniques . The resulting product is highly pure, with a purity greater than 95% as determined by SDS-PAGE .
The biological activity of recombinant glucokinase is measured by its ability to catalyze the phosphorylation of glucose to glucose-6-phosphate. The specific activity of the enzyme is typically greater than 70 units/mg, where one unit is defined as the amount of enzyme that will oxidize 1.0 µmole of glucose to glucose-6-phosphate per minute in the presence of beta-NADP at pH 9.0 and 37°C .
Recombinant glucokinase from E. coli is used in various biochemical and physiological studies, particularly those related to glucose metabolism and regulation. It is also employed in high-throughput screening assays for drug discovery and development, as well as in the study of enzyme kinetics and mechanisms .
For optimal stability, the recombinant glucokinase should be stored at 4°C if it will be used within 2-4 weeks. For longer-term storage, it is recommended to store the enzyme at -20°C, with the addition of a carrier protein such as 0.1% HSA or BSA to prevent degradation. It is important to avoid multiple freeze-thaw cycles to maintain the enzyme’s activity .
