INTRODUCTION

Chymotrypsin is a digestive enzyme that speed up digestion process in the body. It

belonging to a super family of enzymes called serine proteases. Its function to break the
protein down to produce product. In human body, chymotrypsin naturally produced by the
pancreas. However, for some cases it taken as an enzyme supplement to improve health and
digestion and aid in the treatment of various diseases such as asthma, bronchitis, lung
diseases and sinus infection.

Pancreas is a digestive organ that produces chymotrypsin and other digestive enzymes
that required for digestion and absorption of food. The organ is located just below the
stomach in the abdomen. The pancreas secrets pancreatic juice, consisting of enzymes,
water, and electrolytes into the small intestine every day. The enzymes will not digest the
pancreas because they are secreted in an inactive form (as proenzymes). An inhibitor secreted
by the pancreas to ensure that the enzymes are not activated too early. The enzymes become
activated when the pancreatic juice reaches the small intestine and the small intestine is not
digested because it contains a protective mucous lining.

The chemical bonds that hold the food molecules together break down food by the
enzymes secreted by the pancreas. Enzymes lipase secreted to digests fat, enzymes amylases
to break down starch molecules into smaller sugars and protease that breaks protein
molecules into dipeptides and some single amino acids . Other protease enzymes secreted by
the pancreas besides chymotrypsin is include trypsin and carboxypeptidase.

Chymotrypsin is hydrolase type of enzyme. During the breakdown process, it adds a

water molecule. The hydrolysis of peptide bonds of proteins in the small intestine catalyszes
by chymotrypsin. In addition, it also catalyzes the hydrolysis of ester bonds.
HISTORY

In the early 1900s, Vernon proposed that pancreatic preparations could give rise to an
intrinsic activator of its own enzymes.

In 1902, Vernon’s milk-clotting experiments determined there were at least two enzymes
present and that one was more stable than the other. However, this idea was not widely
accepted.

In 1934, Kunitz and Northrop confirmed the presence of an enzyme in addition to trypsin,
naming it chymotrypsin. They were able to crystallize chymotrypsin, as well as the inactive
precursor, chymotrypsinogen.

In 1938, Kunitz isolated different active forms of chymotrypsin, designating them as alpha,
beta, and gamma.

In the early 1940s, Fruton and Bergmann further studied the specificity of chymotrypsin,
reporting on several new substrates.

In 1947, Jacobsen identified additional forms of chymotrypsin, designating them as delta and
pi.

In 1948, Schwert further characterized the molecular weights of chymotrypsin and

chymotrypsinogen.

In 1954, Hartley and Kilby reported the first evidence for the three-step mechanism of
chymotrypsin hydrolyzing amide and ester substrates.

In 1955, Laskowski obtained a second crystalline chymotrypsinogen, naming it

chymotrypsinogen B.

PROPERTIES

Molecular weight : 25.6 kDa

Optimal pH : 7.8 – 8.0

Optimal temperature : 40 – 50 ◦C

Isoelectric point : 8.33 (Chymotrypsin, theoretical)

8.52 (Chymotrypsinogen, theoretical)

Extinction coefficient : 51840 cm-1 M-1 (Theoretical)

Active side : Hisdistine

: Aspartate

: Serine
CHYMOTRYPSIN MECHANISM

Step 1: The side of chain of the residue next to the peptide bond to be cleaved nestles in a
hydrophobic pocket on the enzyme when the polypeptide binds, positioning the
peptide bond for attack. One proton from serine extracts to form an alkoxide ion by
histidine. Polypeptide reacts with the serine ion.

Step 2: In chymotrypsin, the carboxylate R-group of Asp102 forms a hydrogen bond with R
group of His 57. This hydrogen bond compresses and electron density shifts to the
other nitrogen atom in the R-goup of His57 becomes a very strong base. It turn
Ser195 into a strong nucleophile that can attack the substrate by allow His 57 to
deprotonate Ser195.

Oxygen develops a partially negative charge in the oxyanion hole (a pocket in

active side of an enzyme).

Step 3: Collapse of the tetrahedral intermediate, re-formation of a double bond with carbon
which breaks the peptide bond between the carbon and amino acid group happened
due to instability of the negative charge on the substrate carbonyl oxygen. The
 amino leaving group is protonated by His57, facilitating its displacement. The bond
breaks after the oxyanion hole stabilizes the negative charge. The leaving group is
stabilized and the acyl-enzyme is formed.

Step 4: The body metabolized the depart of amine component from the enzyme and it binds
to serine. First stage acylation of enzyme completed. The first product produce.

Step 5: A water molecule is added. The water deprotonated to form a hydroxy group by
Histidine. Hydroxy group and carbon from the carboxy side attaches then
destabilizes the acyl intermediate. The bond is broken.

Step 6: Acid-base catalysis deprotonates an incoming water molecule, generating a strongly

nucleophilic hydroxide ion. A second tetrahedral intermediate generated due to
attack of hydroxide on the ester linkage of the acylenzyme.

Step 7: The second product, a carboxylate anion form after collapse of the tetrahedral
intermediate. The proton goes back to Serine from Histidine.

Step 8: The carboxylic acid is released. The enzyme is reformed to its original shape and
active site to catalyze the next reaction.

GENERAL USE

Generally, chymotrypsin are primary use as a digestive aid and as an anti-inflammatory

agent because it has proteolytic, anti-inflammatory, and antioxidant properties. In some cases,
the presence and amount of chymotrypsin measured for diagnostic purposes as a test of
pancreatic function.

Chymotrypsin along with the other pancreatic enzymes such as lipase and amylase, is
most often used in the treatment of pancreatic insufficiency (not enough of the digestive
enzymes normally secreted by the pancreas into the intestine). Pancreatic insufficiency is
characterized by impaired digestion, malabsorption and passing of undigested food into the
stool, nutrient deficiencies, gas, and abdominal bloating and discomfort. Persons with cystic
fibrosis that is hereditary disease may also suffer pancreatic deficiency. Besides, it may also
occur in those with chronic pancreatitis. Other conditions that, physical injuries,
chemotherapy, and chronic stress could result in chymotrypsin deficiency.
 Without the help of pancreatic enzymes, other molecules such as starch and fat digestion
can be accomplished. However, the protease enzymes including chymotrypsin, trypsin, and
carboxypeptidase are required for proper protein digestion. The development of allergies and
the formation of toxic substances produced by putrefaction, the breakdown of protein
materials by bacteria may occur because of the incomplete digestion of proteins. To keep the
small intestine free from parasites such as bacteria, yeast, protozoa, and intestinal worms,
protease enzymes and other intestinal secretions are required. Pancreatic function asses by a
laboratory analysis of a stool sample along with physical symptoms.

Specifically, chymotrypsin is used to:

 Aid in digestion.
 Treat inflammation and reduce swelling. For example, soft tissue injuries, acute
traumatic injuries, sprains, contusions, hematomas, ecchymoses, infections, edema of the
eyelids and also genitalia, muscle cramps, and sports injuries.
 Treat arthritis and such other autoimmune diseases as lupus, scleroderma, and multiple
sclerosis.
 Treat ulcerations and abscesses.
 Liquefy mucus secretions.
 Treat enterozoic worms and other parasites in the digestive tract.
 Treat cancer. But a controversial use that requires much more scientific study, though
chymotrypsin may be helpful in alleviating effects of radiation treatment or
chemotherapy.
 Treat shingles and acne.
 Decrease effects of sun damage and age sp
PREPARATION

Chymotrypsin is produced from animal such as pig, cows, or bullock pancreas. It can be
taken in three ways that are orally, topically, or by injection. It is commonly taken orally in
tablet form and injection is only for some certain cases. As a tablet, it may be uncoated,
microencapsulated, or even enterically coated. It is coated to prevent digestion in the stomach
so that the enzyme will be released in the small intestine. Other than, coated granules,
powder, capsules, and liquids are the other form of chymotrypsin. As for burns, wounds, and
abscesses, creams and ointments are used to break down proteins and debride dead tissue.
The enzyme preparation should be stored in a tight container with a moisture-proof liner in a
dry, cool place.

Usually chymotrypsin contained not just the enzyme itself but combined with other
enzymes. A typical formulation are chymotrypsin (0.5–1 mg), a plant protease known as
bromelain(25–45 mg), a mixture of many pancreatic enzymes called pancreatin(100 mg),
another plant protease that have same function as chymotrypsin called papain(25–60 mg),
and a pancreatic protease that is trypsin(24 mg). To enhance the activity of the enzyme
supplement, formulations may also include vitamins, herbs, phytochemicals, and other
nutrients.

In the preparation usually in tablet form, the dose required will different on the quantity
(amount in mg) and the quality (activity level) of the enzyme. The dose will depend on the
condition being treated. For oral ingestion and for topical application, the directions on the
bottle or tube label mostly can be followed. As for the enterically coated tables, it should be
swallowed and not chewed or ground up. At least 8 oz of water is taken to take the tablets to
help activate the enzyme. Chymotrypsin usually taken just before, during, or just after meals,
or before going to bed at night to enhance digestion. Improvements in digestion should be
noted within a few hours with proper dosages taken.

Chymotrypsin should be taken on an empty stomach for inflammatory or chronic

conditions, its either one hour before meals or at least two hours after meals. For an
inflammatory condition, some improvement may be noted within three to seven days when
chymotrypsin is taken. Those with chronic conditions such as arthritis may required more
time to notice a change in conditions, it may take three months or more.
SIDE EFFECTS OF THE PREPARATION

Chymotrypsin therapy won’t cause any long-term side effects from if precautions for its use
are followed.

Some people should not take enzyme therapy including:

 People with hereditary clotting disorders such as haemophilia

 People that suffering from coagulation disturbances
 People that just about to or have undergone surgery
 People that on anticoagulant therapy
 People that suffering from protein allergies
 Pregnant women or those breast-feeding

Studies have shown that at recommended doses, after 24–48 hours, enzymes cannot be
detected in blood analysis.

There are temporary side effects that may occur. The effects should be disappear when
therapy is discontinued or dosage is reduced. The effects are:

 Changes in the color, consistency, and odor of the stoo.

 Gastrointestinal disturbances such as flatulence
 A feeling of fullness
 Diarrhea
 Constipation
 Nausea
 Minor allergic reactions such as reddening of the skin.

CONCLUSION

In conclusion, chymotrypsin is important enzyme that needed in protein digestion. Unlike

other substances, the digestion of protein won’t be completed without the protease enzymes,
chymotrypsin and trypsin. People that have problem in the digestion should take the enzyme
therapy and taking the enzyme preparation as a solution. Besides, preparation of
chymotrypsin also used for many other health problems. It shown that chymotrypsin is
important in our life.

REFERENCES
http://www.worthington-biochem.com/CHY/
https://www.slideshare.net/VikramAditya41/chymotrypsin-serine-protease-mechanism

https://commons.wikimedia.org/wiki/File:Oxyanion_hole_stabilizes_the_tetrahedral_interme
diate.png

https://en.wikibooks.org/wiki/Structural_Biochemistry/Enzyme_Catalytic_Mechanism/Protea
ses/Chymotrypsin

https://www.encyclopedia.com/science-and-
technology/biochemistry/biochemistry/chymotrypsin
https://www.rxlist.com/chymotrypsin/supplements.htm
https://vitagene.com/supplements/compounds/chymotrypsin
https://www.drugbank.ca/drugs/DB09375