Introduction Of Recombinant bibasic amino acid endopeptidase (KEX2)

Recombinant bibasic amino acid endopeptidase (Kex2; E.C.3.4.21.61) is a precursor processing enzyme belonging to the subtilis protease family, capable of specifically recognizing and cleaving the carboxyltelopeptide bonds of Lys-Arg and Arg-Arg bibasic amino acids. This enzyme is widely used in the processing of insulin analogue precursors and other enzymatic digestion processes for peptide or protein drugs.
Precursor processing enzymes are a class of proteolytic enzymes that are widely present in living organisms and play a shear processing role in the maturation of many peptides and proteins, and the concept of precursor processing was proposed by two laboratories more than 50 years ago [1,2]. Later, it was gradually discovered that the substrates processed by precursor processing enzymes include not only polypeptide hormones, but also neurotransmitters, growth factors, plasma proteins, viral coat glycoproteins, cell surface receptor proteins, and bacterial exotoxins. In recent years, the understanding of various precursor processing enzymes has gradually deepened, and the genes of various precursor processing enzymes have also been discovered, which has also shown their importance in cell cycle, apoptosis, antiviral drugs and gene regulation.
Our recombinant bibasic amino acid endopeptidase amino acid sequence is derived from Saccharomyces cerevisiae, recombinant expression by Pichia Pichia, theoretical molecular weight is 74kDa, optimal digestion pH is 8.0~10.0, optimal digestion temperature is 37°C, digestion activity is not inhibited by serine protease inhibitors such as PMSF, TPCK, etc., and is stable under pH 5.0~6.0 conditions.
Application of Kex2
Kex2 has great potential for application in the biopharmaceutical industry. At present, there are mainly the following aspects:

1. When designing the protein sequence, the Kex2 cleavage site is introduced, and the expression system is applied to Pichia yeast expression system, which is recognized and digested by the yeast's own Kex2 enzyme in the later secretion and expression process, so as to achieve a large amount of expression of recombinant protein. There are many relevant literatures in China, such as the secretion and expression of recombinant human insulin in Pichia [3], the secretion and expression of recombinant human parathyroid hormone pTH (1-34) in Pichia [4], and the secretion and expression of human epidermal growth factor (hEGF) in Pichia [5].
2. When designing the protein sequence, the Kex2 cleavage site is introduced, the protein is expressed by the Escherichia coli expression system, and the enzyme cleavage is performed with Kex2 in the later preparation process to obtain the target protein. The cutting of amino acid residues in this type of protein usually uses chemical method or enzyme cleavage method, while most of the chemical methods in the more general methods cannot avoid the modification of the target peptide and increase the purification cost, while the enzyme method is more likely to use such as achromatic bacillus enzyme I or trypsin, etc., these enzymes usually recognize an amino acid residue in use, which is highly restrictive to the sequence of the target protein, and may cause problems such as miscleavage, so the production and preparation of enzymes such as Kex2 that recognize several amino acid residues and have high specificity is particularly important. After review, it has been reported that the chimeric protein βGal-117S4HPPTH34 expressed by Kex2 enzyme can be used to cleave, from which recombinant human parathyroid hormone pTH (1-34) has been cleaved [6]. 3. Process digestion in peptide drug production, biological research: enzymatic digestion of proteins and peptide mapping analysis, sequencing, etc.

Our Kex2 product features:
High purity, high specific activity, and host protein residue is less than the limit requirements of biological products.
Good stability: Freeze-dried powder, easy to store and transport.
No animal origin: recombinant production, no exogenous virus contamination, no animal origin raw materials are used in the production process.
3,000L fermentation scale under the GMP quality management system: the batch output scale is large and the batch consistency is high, and the batch output can reach 500-1,000g, which can meet the quality requirements of scientific research customers and the requirements of industrialization customers for product quality, output and GMP conditions during industrialization.
Complete quality documentation: Relevant regulatory support documents can be provided according to customer needs.
Matching Kex2 trace residue detection kits are also available.

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