Trypsin Porcine
E.coli.
Sterile Filtered lyophilized powder.
Description
Recombinant Porcine Trypsin is expressed in E.coli and purified by standard chromatography techniques.
Product Specs
Trypsin (EC3.4.21.4), a member of the serine protease family, cleaves peptides at the C-terminal side of lysine and arginine residues. The hydrolysis rate decreases if an acidic residue is adjacent to the cleavage site and is completely absent if a proline residue is present on the carboxyl side. Trypsin exhibits optimal activity between pH 7 and 9. Additionally, trypsin hydrolyzes ester and amide linkages in synthetic amino acid derivatives such as benzoyl L-arginine ethyl ester (BAEE), p-toluenesulfonyl-L-arginine methyl ester (TAME), tosyl-L-arginine methyl ester, N-α-benzoyl-L-arginine p-nitroanilide (BAPNA), L-lysyl-p-nitroanilide, and benzoyl-L-tyrosine ethyl ester (BTEE). Recombinant trypsin can be inhibited by serine protease inhibitors including TLCK (N-p-tosyl-L-lysine chloromethyl ketone), PMSF (phenylmethanesulfonyl fluoride), benzamidine, soybean trypsin inhibitor, and ovomucoid.
Recombinant Porcine Trypsin is produced in E. coli and purified using standard chromatographic techniques.
Sterile Filtered Lyophilized Powder
The Porcine Trypsin is lyophilized with mannitol as a preservative.
Reconstitute the lyophilized Porcine Trypsin in sterile 1mM HCl or 50mM HAC to a concentration of at least 100 μg/ml. This solution can be further diluted in other aqueous solutions.
Recombinant Porcine Trypsin, while stable at room temperature for one week, should be stored desiccated below -18°C. For long-term storage, adding a carrier protein (0.1% HSA or BSA) is recommended. Avoid repeated freeze-thaw cycles.
4500 USP units per mg of protein
One USP unit of trypsin activity is defined as the amount of enzyme that produces a change in absorbance at 253 nm (ΔA253) of 0.003 per minute in a 3.0 ml reaction volume at pH 7.6 and 25°C using BAEE as the substrate (1 cm light path).
For trypsin digestion, a weight ratio of trypsin to substrate ranging from 1:50 to 1:1000 is recommended.
E.coli.
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DVLEGNEQFINAAKIITHPNFNGNTLDNDIMLIKLSSPATLNSRVATVSLPRSCA
AAGTECLISGWGNTKSSGSSYPSLLQCLKAPVLSDSSCKSSYPGQITGNMICVGF
LEGGKDSCQGDSGGPVVCNGQLQGIVSWGYGCAQKNKPGVYTKVCNYVNWI
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Product Science Overview
Recombinant porcine trypsin is produced by inserting the gene encoding for porcine trypsin into a host organism, such as the yeast Pichia pastoris. This host organism then expresses the trypsin enzyme, which can be harvested and purified for various applications . The amino acid sequence of recombinant trypsin is identical to that of naturally occurring porcine trypsin, ensuring that it retains the same enzymatic properties .
Recombinant porcine trypsin is widely used in the biotechnology and pharmaceutical industries. Some of its key applications include:
- Cell Culture: It is used to detach adherent cells from culture vessels during the passaging process. This is essential for maintaining and expanding cell cultures in research and production settings .
- Protein Production: In the manufacture of recombinant proteins, such as insulin, trypsin is used as a protein-cleaving reagent during the downstream processing steps .
- Diagnostic Assays: Trypsin is used in various diagnostic assays and research applications to study protein structure and function.
The recombinant production of porcine trypsin offers several advantages over traditional extraction methods:
- Purity: Recombinant trypsin is free from other proteases, such as chymotrypsin, which can be present in naturally derived trypsin .
- Consistency: The production process is highly controlled, ensuring consistent enzyme activity and quality.
- Safety: Recombinant trypsin is free from contaminants and adventitious agents that might be present in animal-derived trypsin, reducing the risk of contamination in sensitive applications .
The use of porcine trypsin in the manufacture of human biological medicinal products is subject to regulatory guidelines to ensure safety and efficacy. These guidelines cover aspects such as the source of the trypsin, testing for adventitious agents, and validation of the manufacturing process . The European Medicines Agency (EMA) has published guidelines on the use of porcine trypsin, highlighting the importance of quality control and risk assessment in its production and application .
