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Description
Unwanted C-terminal basic amino acids — lysine, arginine or ornithine — can block protein sequencing, interfere with peptide map interpretation, and leave charged tails on recombinant proteins and processed peptides. Removing them cleanly and reproducibly with a well-characterized exopeptidase is a routine first step in protein analysis and downstream processing.
Carboxypeptidase B from porcine pancreas (EC 3.4.12.3) is a zinc-containing metallocarboxypeptidase that selectively hydrolyzes peptide bonds at the C-terminal basic amino acids lysine, arginine and ornithine:
Peptidyl-L-lysine + H2O → Peptide + L-Lysine
Also known as protaminase or peptidyl-L-lysine [L-arginine][L-arginine][L-arginine] hydrolase, with a molecular weight of 34,300 Da.
What is the source and form of this product?
This Carboxypeptidase B is derived from porcine pancreas and supplied as a lyophilized powder.
How should Carboxypeptidase B lyophilized powder be stored and reconstituted?
Store the lyophilized powder at -20°C. Reconstitute by dissolving in distilled water or a dilute buffer.
What is the specific activity and protein content of this Carboxypeptidase B?
The specific activity is approximately 50 U/mg protein, with a protein content of approximately 80% as determined by the biuret method.
How is Carboxypeptidase B activity defined?
One unit of Carboxypeptidase B hydrolyzes 1 µmol of hippuryl-L-arginine per minute at 25°C and pH 7.65.
How is Carboxypeptidase B activity measured?
Activity is measured by the increase in absorbance at 254 nm resulting from the hydrolysis of hippuryl-L-arginine. The initial linear rate (ΔA254/min) is used together with the hippuric acid extinction coefficient of 0.349 to calculate units per mg enzyme.
Which amino acids does Carboxypeptidase B cleave?
Carboxypeptidase B selectively hydrolyzes the C-terminal basic amino acids lysine, arginine and ornithine. Example reaction: Peptidyl-L-lysine + H2O → Peptide + L-Lysine.
Is Carboxypeptidase B a metalloenzyme?
Yes. Carboxypeptidase B is a zinc-containing metalloenzyme (metallocarboxypeptidase); one atom of zinc is required for catalytic activity.
For research use only. Not for use in diagnostic or therapeutic procedures.
