Chemical Reagent

High Purity Recombinant Carboxypeptidase B is Used for Producing Recombinant Polypeptide

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Description

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Pancreatic or tissue Carboxypeptidase B (CPB), a key enzyme involved in insulin conversion and highly specific for excising C-terminal Lys and Arg residues from peptides and proteins, was expressed at high level and purified from a recombinant Pichia pastoris strain. A cDNA containing the porcine pancreatic pro-Carboxypeptidase B (proCPB) fused to the Saccharomyces cerevisiae alpha factor secretion signal was cloned into the pPIC3K vector under control of P. pastoris AOX1 promoter. After 72 h of growth on methanol, proCPB accumulated until 320 mg L?1, representing 70% of total proteins in culture supernatant. A single stepwise ion exchange purification process with Q-Sepharose at increasing concentrations of ammonium acetate allowed recovery of 65% proCPB in a single fraction.? Recombinant Carboxypeptidase B is expressed in E.Coli?and purified by high pressure liquid chromatography. There is no trace of other enzyme (such as carboxypeptidase A and chymotrypsin) activity. No protease inhibitors such as PMSF are present in the preparation.

Physical form:Lyophilized from 20 mM Tris, pH 8.0 + 50 mM NaCl.

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Advantages

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Animal origin free:YaxinBio recombinant carboxypeptidase B belongs to the AOF level 3, eliminate the risk of virus presence, or any other potential adventitious agents found in animal-derived carboxypeptitase B

Stability:A sterile recombinant carboxypeptidase B lyophilized eliminates the risk of contamination and decreases the chances of activity loss in the process of transport and storage.

High purity:

1) Recombinant carboxypeptidase B provides increased specific?activity?and?eliminates contaminating proteases activities found in extracted enzymes with lower purity?level.

2) No other contaminating proteases such as chymotrypsin and carboxypeptidase A.

3)Less than 10ppm of recombinant trypsin.

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Reconstitution


Centrifuge the vial prior to opening. Reconstitute in water to a concentration of 0.1-1.0 mg/mL. Do not vortex. This solution can be stored at 2-8°C for up to 1 week. For extended storage, it is recommended to further dilute in a buffer containing a carrier protein (example 0.1% BSA) and store in working aliquots at -20°C to -80°C.
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Biochem/physiol Actions


Proteases (also called Proteolytic Enzymes, Peptidases, or Proteinases) are enzymes that hydrolyze the amide bonds within proteins or peptides. Carboxypeptidase-B sequentially cleaves C terminal K and R residues. Recombinant rat Carboxypeptidase-B is a 35.1 kDa protein consisting of 307 amino acids.