Prepared by:

Ms. Hetal [Link]

 Introduction
An ENZYME is a PROTEIN that functions as a biocatalyst
to SPEED UP a CHEMICAL REACTION in the body.

Charateristics:
1. The basic function of an enzyme is to increase the rate of
reaction. Most cellular reactions occur about a million times
faster than they would in the absence of an enzyme.
2. Most enzymes are specific with only one reactant.(called a
substrate)
3. The enzymes are regulated from a state of low activity to
high activity and vice-versa.
 Introduction contd...
Enzymes can be kept in three categories:
1. Metabolic enzymes
2. Digestive enzymes
3. Food enzymes

Factors affecting Enzyme activity:

• Most enzymes act best at temperatures between 35 and 40 ºC.
Temperatures above 65 ºC , especially in the presence of moisture,
destroy them, whereas their activity is negligible at 0ºC.
• Certain heavy metals, formaldehyde and free iodine retard the
enzyme activity.
• Their activity is affected by the pH of the medium in which they act
or by the presence of other substances in the medium.
• They are highly selective in their action.
 Introduction contd...
• Enzymes are either obtained from plants or they are
manufactured by the pancreas.

• Pancreatic enzymes are animal based and only function in our

small intestine. If pancreatic enzymes are taken with food ,
they will be destroyed by the acids in our stomach therefore,
they are not nearly as effective as plant enzymes.

• Plant enzymes are much more effective because they begin

pre-digestion in our mouth, they are destroyed by the acids in
our stomach and they function in both an acid and in alkaline
environment.
Classification
Enzymes are also classified on the basis of site of action :
1. Exoenzymes ( extracellular enzymes)
• secrerted outside the cell
• used in digestion
• e.g. Proteoses, amylase, lipase

2. Endoenzymes (Intracellualar enzymes)

• act inside the cell
• responsible for synthesis of cell component, food reseves ,
liberation of energy.
• e.g. Synthetase, isomerase
Other Classification:
1. The Amylolytic enzymes OR carbohydrates – Diastase,
Amylase, Invertase, Sucrase, ptyalin, Amylopsin, Malt
diastase.

2. The esterases – Lipase , Pectase, Steapsin, Urease.

3. The proteolytic enzymes – Pepsin, Trypsin, Papain , Rennin,

Erepsin

4. The oxidizing enzymes – Peroxidases, Thrombin, Zymase

 Properties of Enzyme

1. Sensitive to heat and are denatured by excess heat or cold. their

active site becomes permanently wrapped, thus the enzymes is
unable to form the enzyme substrate complex.

2. Enzymes are created in cells but are capable of functioning

outside of the cell.

3. Enzymes are sensitive to pH, the rate at which they can conduct
reactions is dependent upon the pH of where the action is taking
placed. For Example, pepsin in stomach has an optimum pH of
about 2, whereas salivary amylaze has an optimum pH of about
7.
4. Enzymes reusable and some enzymes are capable of
catalysing many hundreds of thousands of reactions.

5. Enzymes will only catalyse one reaction, for example ,

Invertase will only produce Glucose and Fructose, when a
glucose solution is passed over beads of enzyme.

6. Enzymes are capable of working in reverse. This acts as a cut

off point for the amount of product being produced. If there
are excess reactants , the reaction will keep going and be
reversed, so that there is no overload or build up of product.
 Uses of Enzyme
1. The uses of important enzymes in medicine include killing disease-
causing micro-organisms, prompting wound healing, and
diagnosing certain diseases.
2. Pharmaceutical enzyme applications generally require small
quantities of highly purified enzymes.
3. Enzymes can be used to detect and measure amounts of glucose in
blood. Amount of glucose in blood and urine is an indicator for
diagnosis of diabetes. Detected by using enzyme glucose oxidase.
4. Can be administered individually or along with other drugs and/or
treatments.
 Enzyme supplements for enzyme deficiencies.
 Collagenase treats skin ulcer.
 Asparaginase used to treat leukemia.
 Streptokinase administered to patients immediately after heart
attacks
 Enzymes to be covered
1. Diastase
2. Papain
3. Pepsin
4. Trypsin
5. Pancreatin
6. Bromalein
7. Ficin
8. Penicillinase
9. Hyalluronidase
10. Streptokinase
11. Urokinase
 1. Diastase
Synonym: Amylase

Biological source:
• It is a yellowish white, amorphous powder obtained from an
infusion of malt (known as malt-diastase) containing malt
splitting enzyme α -glucosidase

• It is amylolytic enzyme present in digestive tract of animals

known as animal - diastase.

• If present in pancreas it is pancreatic diastase (amylopsin) and

• If present in saliva it is salivary diastase (ptyalin ).

Description :
Colour- yellowish white
Odour – characteristic
Nature – amorphous powder
Solubility – forms a colloidal solution with water, it precipitates in
alcohol
Extra features – thermolabile and denatures at a temperature above
45º C and a pH less than 4. Best active at temperature 35-40ºC
and pH 6-7.

Uses :
• As a digestant,
• Used in production of pre-digested starchy foods and also for the
conversion of starch to fermentable sugars in fermentation.
• In brewing industries.
 Chemical tests
• Principle: Amylase digests starch to form monosaccharides
and starch + I 2 - Blue colour, so, in presence of amylase, I 2
will not give blue colour.
 2. Papain
Biological source:
  Papain consists of the dried latex of the green fruits of Carica papaya
Linn. Family: Caricaceae.
Preparation:
• Vertical incisions are made in the skin of the immature fruit while it
is still hanging on the tree.
• The latex is collected in aluminium trays and to the latex potassium
metabisulphate is added in a proportion of 5 gm per kg of latex.
• Extraneous matters are cleaned by passing it through sieve and the
latex is dried in vacuum at a temperature of 55-60 ºC to obtain
papain.
• Incisions and collection are made at weakly intervals. Fruit exudes
the latex (between 5 and 10 A.M.).
• It is purified by dissolving in water and precipitating with alcohol.
Description :
Colour- light brown or white
Odour – characteristic
Taste – characteristic
Solubility –solution in water and glycerine
Best active–pH of 5-6.

Chemical Name – mixture of papain and chymopapain

Identification:
1. It decolorizes the aqueous potassium permanganate solution.
[Link] causes curdling of milk.(Proteolytic effect).
Constituents :
• It contains several enzymes that include one or more
proteolytic enzymes.
• Papain, a coagulating rennet-like enzyme which acts upon
the casein of milk;
• an amylolytic enzyme;
• a clotting enzyme similar to peptase;
• It is quite apparent that more than one proteolytic enzyme is
present because a single sample of papain will yield variable
results depending upon the protein used in the substrate. The
best grade digests 300 times it’s weight of egg albumin
Uses :
•  Papain is used as a digestant for proteins.
• As an anti inflammatory agent
• as an ingredient in cleaning solutions for soft contact lenses.
• Papain is used extensively for tenderizing beef.
• It is used in meat packing industries.
• It is used in relieving symptoms of episiotomy (An episiotomy
also known as perineotomy, is a surgically planned incision on
the perineum and the posterior vaginal wall during second
stage of labour).

Chymopapain is a non-proteolytic enzyme obtained from the

latex of Carica papaya Linn. V Family: Caricaceae. It is
sulfhydryl enzyme.
 3. Pepsin
Biological source:
  It is a proteolytic enzyme obtained from the mucous membrane of the
fresh stomach of various animals like pig, sheep. The commonly
used species of pig is Sus scrofa Linn. var. domesticus gray.
Family: Suidae.
Preparation:
• The mucous membrane is separated from the stomach by stripping
or scapped off and it is placed in acidified water for autolysis at
37 ºC for 2 hours.
• The liquid obtained after autolysis consist of both pepsin and
peptone.
• It is then filtered and sodium or ammonium salts are added to the
liquid till it is half saturated.
• At this point only the pepsin separates out and the peptone remains
in the solution.
• The precipitates are collected and subjected to dialysis for the
separation of salts,.
• Remaining amount of pepsin if any in the aqueous solution is
precipitated by the addition of alcohol into it.
• The pepsin is collected and dried at low temperature.

Description:
Colour- Pale yellow
Odour – odorless or very faint odour
Tatse- slightly bitter
Solubility –soluble in water, dilute acids and physiological salts (NaCl)
solution
Best active– at a temperature of 40 ºC with pH of 2-4 .
• Pepsin is unstable above pH 6.
• The enzyme gets denatured at a temperature of 70ºC and in the
presence of alcohol and sodium chloride.
• Pepsin can be stored for 1-2 years at 2-8 ºC.

Uses :
• Used in the deficiency of gastric secretion.
• Pepsin is also used in the laboratory analysis of various proteins.
• In the preparation of cheese and other protein containing foods.
 4. Trypsin
Biological source:
  It is a proteolytic enzyme obtained produced by Ox pancreas, Bos
taurus. Family: Bovidae

It is one of the three principal digestive proteinases which along with

the other proteinases like pepsin and chymotrypsin break the
dietary protein molecules to their amino acids and peptide
component.
Production: Trypsin is produced by pancreas in the form of
trypsinogen. Trypsin is then transported to the small intestine,
where the proteins are cleaved into polypeptides and amino acids.
As trypsin is an autocatalytic enzyme , it by itself catalyses the
conversion of trypsinogen to trypsin.
Another enzyme enterokinase is also required in small amount to
catalyze the initial reaction of trypsinogen to trypsin.
 Properties :
 It is yellowish white powder without any odour.
 It is soluble in water, insoluble in organic solvent.
 Trypsin has maximum enzymatic activity at pH 8.
 It should be freshly prepared.
Storage:
 It should be stored at very cold temperatures (2 to 8 C) to prevent
autolysis (Self -cleavage).
 Autolysis may also be prevented by storage of trypsin at pH 3.
when the pH is adjusted back to pH 8 activity returns.
Uses:
 In a tissue culture lab ,it is used to re-suspend cells adherent to the
petridish wall during the process of harvesting cells.
 It is also used to harvest corn and oats.
 Trypsin is vital in a cow’s diet , without it they would not be able to
digest the grass.
 5. Pancreatin
Biological source:It is a digestive enzyme extracted from the pancreas
of certain animals like hog Sus Scrofa (Suidae) or Ox, Bus taurus
(Bovidae) .

Description:
Colour- cream colored amorphous powder
Odour – faint, characteristic but no offensive
Best active– in neutral or faint alkaline solutions. More than traces of
mineral acid or large amounts of alkali hydroxides make it inert and
alkali carbonates inhibits its action.

Constituents : it contains three enzymes amylopsin (amylase) , trypsin

and strpsin (lipase).
Uses :
• It has two important functions in the body – digestion of foods and
routine cancer eradication.
• It is an effective enzyme supplement for replacing missing
pancreatic enzymes used in a number of essential body procedures.
• Enteric coated granules of pancreatin are used in treating infants
with celiac diseases and related pancreatic deficiencies.
• It has also been claimed to help with food allergies, celiac disease,
autoimmune disease, cancer and weight loss.
 6. Bromalein
Biological source:
  It is a mixture of proteolytic enzymes obtained from the stem and
ripen fruits of pineapple plant Ananas comosus Family:
Bromeliaceae

Preparation : It is isolated from pineapple juice by precipitation

with acetone and also with ammonium sulphate

Description:
Odour - Odorless to slightly putrid
Color – light brown
Taste – irritating
Solubility: Soluble in water, insoluble in organic solvents like
ether, chloroform and alcohol
Constituents : It is a collection of enzymes and other compounds. It
is a mixture of sulfur containing protein digesting enzymes,
called proteolytic enzymes or proteases.
It also contains several other substances in minor quantities ,
including peroxidase, acid phosphatase, protease inhibitors and
calcium.
Use:
• It is an effective fibrinolytic agent, it inhibits platelet
aggregation and seems to have both direct as well as indirect
action involving other enzyme systems in exerting its anti
inflammatory effect.
• In treatment of soft tissue inflammation, edema due to surgery
and injury.
• It can modify the permeability of organs and tissues to different
drugs.
• It is useful in the prevention and treatment of thrombosis.
• It may even be useful in the treatment of AIDS to stop the spread
of HIV.
• It has no major side effects except the possible allergic reactions.
• In food Industries – alcohol and beer industries, hydrolisation of
proteins, meat processing, baking industries, pet food, health
food.
• It is also used in cosmetic and textile industries.
 7. Ficin
Biological source: It consists of nearly ripe and sun dried succulent
fruits of the plant Ficus carica family Moraccae.
 Characters:
The optimum pH of ficin depends upon the substrate and its
concentrations. Generally the optimum pH is between 5-8, although
ficin keeps its activity over the range of pH 4-9 at 60 ºC.
Though the optimum temperature of ficin is 45-55ºC . It is effective in
temperatures between 15-60 ºC.
It is obtained as a white to yellow microgranular powder .
The moisture content should not exceed 6 %.
Uses : In food Industries – alcohol and beer industries, hydrolisation of
proteins, meat processing, baking industries, pet food, health food.
Pharmaceutical Industries – contact lens cleaning, cancer treatment,
anti arthritis, digestive aid.
 9. Hyalurodinase
Synonyms : Spreading factor, hyalase
Biological source:
It is an enzyme product prepared from mammalian testes which
shows the capability of hydrolysing hyaluronic acid like
mucopolysaccharides. Skin is considered as the largest store of
hyaluronidase in the body.
Characteristics :
• Hyalurodinase for injection consists of NMT 0.25 µg of
tyrosine for each USP hyaluronidase unit.
• Due to its action on hyaluronic acid , it promotes diffusion and
hastens absorption of subcutaneous infusions.
• It depolymerises and catalyses hyaluronic acid and similar
hexosamine containing polysaccharides.
Constituents :
• Hyalurodinase is a group of enzymes such as 4-lycanohydrolase,
hyaluronate 3-glycanohydrolase and hyaluronate lyase.
• They are mucopeptides composed of alternating N-
acetylglucosamine and glucuronic acid residues.
• Hyalurodinase catalyse the breakdown of Hyaluronic acid.

Uses :
• Hyalurodinase for injection is used in the conditions of
hypodermolysis.
• It is used as a spreading and diffusing agent.
• It promotes diffusion, absorption and reabsorption.
 10. Streptokinase
Synonym : Plasmokinase

Biological source:
It is a purified bacterial protein produced from the strains of
group C β-haemolytic Staphylococcus griseus.
 
Characterisics:
• It is available as a sterile, friable solid or white powder.
• It is soluble in water with maximum activity at pH 7, the
solution at higher concentration is stable for 6 hours at 4 ºC,
otherwise dilute solutions are unstable.
Constituents :
It is the purified bacterial protein with about 484 amino acid
residues.

Uses :
• It is the first available agent for dissolving blood clots.
• It is used in the treatment of pulmonary embolism, venous
and arterial thrombosis and coronary artery thrombosis.
• It is also sometimes administered along with the heparin to
counteract paradixical increase in local thrombin.
 11. Urokinase
Biological source:
  It is a serine protease enzyme isolated from human urine and from
human kidney cells by tissue cultures or by recombinant DNA
technology.

Preparation:
• It is fibrinolytic enzyme produced by recombinant DNA using
genetically manipulated [Link] cells.
• It is produced firstly as prourokinase q.v. And then converted to
active form by plasmin or kallikrein.
• Urokinase used medicinally is also purified directly from human
urine.
• It binnds to a range adsorbents such as silica gel or kaolin which
can be use to initially concentrate and purify the product.
• It can be further purified by precipitation with sodium chloride
or ethanol or by chromatography.
• Human urokinase needs sterile filtration, a septic filling and
freeze drying.

Characteristics :
• It is lyophilised white powder, soluble in water.
• It is an activator of endogenous fibrinolytic system, which
converts plasminogen to plasmin and degrades fibrinogen, fibrin
clots and other plasma protein.
Constituents :
• Urokinase enzymes are serine proteases that occur as a single
low molecular weight and double high molecular weight
polypeptide chain forms.
• They differ in molecular weight considerably.
• A single chain is produced by recombinant DNA technique.
Uses :
• It is used in the treatment of pulmonary embolism, coronary
artery thrombosis and for restoring the potency of intravenous
catheters.
• It is used to dissolve (lyse) fibrin or blood clots in anterior
chamber of eye and in acute massive pulmonary emboli. As it is
derived from human source, it is less antigenic than enzymes
with similar actions like streptokinase.
 Questions....
1. Give the biological source, method of preparation and uses of
Papain.
2. Write a short note on Pepsin.
3. Describe biological source, preparation, chemical tests for
identification and uses of Papain.
4. Describe biological sources, preparation, tests for identification and
uses of Diastase.
5. Give sources, method of preparation, properties and therapeutic
Importance of Diastase enzyme.
6. Enumerate any one drug with its biological source belonging to
proteolytic and lipolytic type enzymes from the syllabus.
7. Write in chemical tests for papain and amylase .
8. Give sources, method of preparation, properties and therapeutic
importance of Papain OR Pepsin enzyme.
 Questions....
10. .Write a note on Diastase enzyme
11. Give the pharmaceutical uses of enzymes.
12. Write biological source and the method of preparation of Papain.
13. Write a short note on Pepsin and Diastase.
14. Write down only the method of preparations of Papain and
Pepsin.
15. Write a short note any one drug belonging to Carbolytic,
proteolytic and lipolytic type enzymes from the syllabus.