Trypsin is a serine protease enzyme that plays a crucial role in the digestion of proteins. It is produced in the pancreas as an inactive precursor called trypsinogen, which is then secreted into the small intestine. Once in the small intestine, trypsinogen is activated to trypsin by the enzyme enteropeptidase (also called enterokinase).
Trypsin functions by cleaving peptide bonds in proteins, specifically those on the carboxyl side of the amino acids lysine and arginine. This proteolytic activity aids in breaking down proteins into smaller peptides and individual amino acids, which can then be absorbed and utilized by the body for various metabolic functions.
In addition to its physiological role in digestion, trypsin is widely used in research and biotechnology applications, such as:
- Cell culture: It is commonly used for detaching adherent cells from the culture vessels for subculturing, cell counting, or downstream experimental applications. In this context, trypsin cleaves the proteins involved in cell adhesion, allowing the cells to be removed from the surface. Trypsin is often used in combination with EDTA, which chelates divalent metal ions and further weakens cell-substrate interactions.
- Protein digestion for mass spectrometry: Trypsin is employed in proteomics research to digest proteins into smaller peptides before their analysis by mass spectrometry. The specific cleavage pattern generated by trypsin allows for the identification of proteins and the determination of their amino acid sequences.
- Tissue dissociation: Trypsin can be used to dissociate tissues into single cells for various experimental purposes, such as flow cytometry or the establishment of primary cell cultures.
It is essential to control the concentration and incubation time when using trypsin, as excessive or prolonged exposure to the enzyme can damage cells or result in unwanted protein degradation.
