Trypsin (EC 3.4.21.

4) is a serine protease from

the PA clan superfamily, found in the
digestive system of many vertebrates, where it
hydrolyses proteins.
Trypsin is produced in the pancreas as the
inactive proenzyme trypsinogen.
Trypsin cleaves peptide chains mainly at the
carboxyl side of the amino acids lysine or
arginine, except when either is followed by
proline. It is used for numerous biotechnological
processes. The process is commonly referred to
as trypsin proteolysis or trypsinisation, and
proteins that have been digested/treated with
trypsin are said to have been trypsinized.
 FUNCTION
 trypsin catalyzes the hydrolysis of
peptide bonds, breaking down proteins
into smaller peptides.
 The peptide products are then further
hydrolyzed into amino acids via other
proteases, rendering them available for
absorption into the blood stream.
 Tryptic digestion is a necessary step in
protein absorption as proteins are
generally too large to be absorbed through
the lining of the small intestine.
When the pancreas is stimulated by cholecystokinin, it is then secreted into the
first part of the small intestine (the duodenum) via the pancreatic duct.
Once in the small intestine, the enzyme enteropeptidase activates it into trypsin
by proteolytic cleavage. Auto catalysis can happen with trypsin with
trypsinogen as the substrate. This activation mechanism is common for most
serine proteases, and serves to prevent autodegradation of the pancreas
 MECHANISMS
 These enzymes contain a catalytic triad consisting of
histidine-57, aspartate-102, and serine-195.
 These three residues form a charge relay that serves
to make the active site serine nucleophilic.
 This is achieved by modifying the electrostatic
environment of the serine. The enzymatic reaction that
trypsin catalyzes is thermodynamically favorable but
requires significant activation energy (it is "kinetically
unfavorable").
 trypsin contains an "oxyanion hole" formed by the
backbone amide hydrogen atoms of Gly-193 and
Ser-195, which serves to stabilize the developing
negative charge on the carbonyl oxygen atom of the
cleaved amides.
 The aspartate residue (Asp 189) located in
the catalytic pocket (S1) of trypsin is
responsible for attracting and stabilizing
positively charged lysine and/or arginine,
and is, thus, responsible for the specificity of
the enzyme. This means that trypsin
predominantly cleaves proteins at the
carboxyl side (or "C-terminal side") of the
amino acids lysine and arginine except when
either is bound to a C-terminal proline.,[5]
although large-scale mass spectrometry data
suggest cleavage occurs even with proline.
 Trypsin is considered an endopeptidase, i.e.,
the cleavage occurs within the polypeptide
chain rather than at the terminal amino acids
located at the ends of polypeptides.
A schematic representation of trypsin interacting with a peptide substrate is shown. The
catalytic residues (His57, Asp 102 and Ser195, yellow) and the enzyme residues that
contact substrate residues are shown (blue). The positively charged arginine side chain
at position P1 of the substrate is attracted by the negatively charged aspartate 189
located at the bottom of the S1 specificity pocket. This interaction as well as five
enzyme-substrate hydrogen bonds at positions P1 and P3 and glycine 193 help to
position the scissile peptide bond (red) for the nucleophilic attack by the polarized
hydroxyl group of Ser 195 (red arrow).