Recombinant   Carboxypeptidase   B   with   High   Purity   is  Used  for  producing  Recombinant   Polypeptide

Recombinant rat Carboxypeptidase B

Description

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, procarboxypeptidase B 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% procarboxypeptidase B 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.

Advantages

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.

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.
 

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.