How much do you know about the application and preparation method of carboxypeptidase B?

Release time:

2021-04-30


Carboxypeptidase B (carboxypeptidase B, CPB for short) is a zinc-containing pancreatic exopeptidase. CPB contains 307 amino acids and has a molecular weight of 35 ku. Because CPB specifically hydrolyzes the basic amino acids (arginine, lysine or ornithine) at the C-terminus of the peptide chain, it can be used for sequence analysis of proteins and peptides, especially to determine the carboxy-terminal amino acid.

Uses of carboxypeptidase B

1. For the hydrolysis of protein C-terminal basic amino acids (lysine, arginine, histidine);

2. For the identification of protein C-terminal amino acids; remove protein C-terminal basic amino acids, etc.

3. Mainly for the production of recombinant insulin, calcitonin and other tool enzymes, here refers to the industrial production of insulin as an industrial enzyme.

4. For the determination of the C-terminal amino acid of protein

5. Used in domestic scientific research units, reagent companies, biotechnology companies, pharmaceutical companies, etc., such as major foreign reagent companies, pharmaceutical factories and biotech companies.

Preparation method of carboxypeptidase B

The current preparation methods for preparing active carboxypeptidase B can be divided into three categories. One is to extract and activate it from animal pancreas. The drawback of this method is that the obtained CPB may contain other infectious substances (such as viruses or other harmful biologically active components) and limit the application of carboxypeptidase B;

 

The second is that the inactive procarboxypeptidase B (proCPB) is expressed by the original or eukaryotic [such as Pichia pasforis], and then proCPB is digested with trypsin to obtain the activity Carboxypeptidase B. The disadvantage of this method is that the expressed proCPB must be "activated" by trypsin to obtain active CPB, and it must be purified by removing trypsin and active peptides;

 

The third is that it is expressed as a fusion protein in bacteria or yeast (such as β-lactosidase-proCPB fusion protein expression), but it still needs to remove the fusion protein to obtain active CPB.

Desheng’s carboxypeptidase enzyme preparations are freeze-dried powder with purity ≥90% (SDS-PAGE detection). The storage conditions are: freeze-dried powders can be stored at -20°C or 4°C for a long time. Note that the company’s enzyme preparations Carboxypeptidase B is used as a raw material for scientific research experiments or kits. It cannot be used in human trials, nor can it be used as a medicine. Those in need are welcome to order and consult.