Enzymes

Pharmacognosist59@yahoo.com

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 What Are Enzymes?

• Act as Catalyst to
accelerates a
reaction
• Enzymes are
Proteins in nature
• Not permanently
changed in the
process

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 Enzymes
• Are specific for
what they will
catalyze
• Are Reusable
• End in –ase
-Sucrase
-Lactase
-Maltase

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 Enzyme-Substrate Complex
The substance
(reactant) an
enzyme acts on
is the substrate

Joins
Substrate Enzyme

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 Active Site
• A restricted region of an enzyme
molecule which binds to the substrate.
Active
Site

Substrate
Enzyme

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 Induced Fit
• A change in the
shape of an
enzyme’s active
site
• Induced by the
substrate

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 Induced Fit
• A change in the configuration of an
enzyme’s active site
• Induced by the substrate.

Active Site
substrate
Enzyme

induced fit

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What Affects Enzyme Activity?

• Three factors:
1. Environmental Conditions

2. Cofactors and Coenzymes

3. Enzyme Inhibitors

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 1. Environmental Conditions
1. Extreme Temperature are the most
dangerous
- high temps may denature (unfold)
the enzyme.

2. pH (most like 6 - 8 pH near

neutral)
3. Ionic concentration (salt ions)

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 2. Cofactors and Coenzymes
• Inorganic substances (zinc, iron) and
vitamins (respectively) are sometimes need
for proper enzymatic activity.

• Example:
Iron must be present in the quaternary
structure - hemoglobin in order for it to
pick up oxygen.

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 Two examples of Enzyme
Inhibitors
a. Competitive inhibitors: are
chemicals that resemble an
enzyme’s normal substrate and
compete with it for the active
site.
Substrate
Enzyme
Competitive inhibitor

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 Inhibitors
b. Noncompetitive inhibitors:
Inhibitors that do not enter the
active site, but bind to another part
of the enzyme causing the enzyme to
change its shape, which in turn
alters the active site.
Substrate Noncompetitive
Enzyme Inhibitor
active site
altered
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Classification
• There are several methods for classification
of enzymes, few of them are:

1. Classification on the basis of reaction

they control.
2. Classification on the basis of site of
action.
3. Classification on the basis of nature of
substrate
Enzyme obtained from Plants or
(Phytoenzymes)

Phytoenzymes includes:

1. Papain
2. Bromelain
3. Malt extract
Enzyme obtained from Animals

1. Rennin
2. Pepsin
3. Pencreatin
4. Pencrealipase
5. Urokinase
6. Asparaginase
7. Hyaluronidase
Phytoenzymes
PAPAIN
Papain is a substance which contains a mixture
of proteolytic enzymes found in the unripe
fruit of papya tree.
Source
• Papain is dried and purified latex or juice of the
fruit of
Carica papya
• Family Caricaceae

Habitat;
• Papya tree is indigenous to tropical America. It is
cultivated in srilanka, Tanzania, pakistan and india
Isolation or collection

• Papain is isolated from an unripe but fully grown

fruit.
• Fruit is cut little deep on 4 sides.
• Latex or juice flows freely for few seconds, soon
coagulates.
• The coagulated Papain is dried.
• This Papain is in crude form.
• The crude Papain is purified.
Chemical constituents

Papain contains several enzymes such as;

• Peptidase: It converts proteins in dipeptides and
polypeptides.
• An amylolytic enzyme: Which convert
carbohydrates into simple units.
• A pectase like enzyme which clots the blood.
• A rennin like enzyme, which act on the milk protein
called casein to coagulate it.
Uses:
• anti-inflammatory
– meat tenderiser
• breaks down tough meat fibres
• long traditional use in America
– teeth whitening agent
• In some toothpastes
– contact lens cleaner
– wound debriding
– digestive aid
Bromelain

• Bromelaine is obtained from the fruit and stem of

Plant called pineapple.

• Botanical source is Ananas sativus.

• It blong from the family Bromeliaceae.
Types:
• There are two types of bromelaine.
i) Stem bromelaine:
This enzyme is found in the
juice of pineapple’s stem.
• This enzyme is obtained from the juice by
precipitation with acetone. It is a basic protein in
nature.
Fruit bromelaine:
This enzyme is found in the
juice of pineapple’s fruit.
• It is acidic in nature.
• It also contains carbohydrate that cannot be
removed by purification procedure.
Description:
• It is a amorphous powder having faint
characteristic odor.
• It is partially soluble in water while insoluble in
alcohols and acetone.
• Uses:
• It is used as adjunctive therapy for reduction of
inflammation and edema.
• It is used to accelerate tissue repair especially
after episiotomy.
• It is used in tenderizing meat.
• It is used in the leather industries.
Malt extract
• Barley is the dried grain of on or more varieties of
Hordeum vulgare
• Family Gramineae.

• Barley is grown throughout the world wherever the

climate is favorable.
Malt
• Malt or malted barely is dried , artificially
germinated barley grain.
• To prepare malt barely grain are kept wet with
water in a warm room and allow to germinate until
the caulicle protrudes.
• The grain is quickly dried.
• The enzyme diastase in the moist warm
grains convert the starch to maltose,
thereby stimulating the embryo to growth.
• The embryo is killed when the grain is dried.
• Dry malt resembles barley but is more crisp ,
has an agreeable odor and has a sweet taste.
Malt extract
• Malt extract is the product obtained by extracting
the malt, the partially and artificially germinated
grain of one or more varieties of Hordeum vulgare
• Family Gramineae.
• The malt is infused with water at 60oC, and the
expressed liquid is concentrated at a temperature
not exceeding 60oC.
Constitutes:
• It contain dextrin, maltose, a small amount
of glucose, and amylolytic enzymes.
• It convert not less than 5 times its weight
of starch into water soluble sugars.
Uses and dose.
• Malt extract is used as easily digested
nutritive and as a aid in digestion of starch.
• The usual dose is 15 g.

• Many commercial extracts of malt do not

• Contain diastase ,which is destroyed by the
heat used for the preparation. Such product
should not confused with this product.
• They are used as bulk laxatives.
Animal Enzymes
1. Rennin:
Rennin is a coagulating enzyme present in the mucous
membrane of the stomach of mammals.
Source:
• Rennin is obtained from the glandular layer
of the stomach of calf
Bos taurus.
Family: Bovidae.
Preparation:
• The glandular layer of the calf’s stomach is
cut into small pieces.
• These pieces are treated with 0.5% sodium
chloride. The solution is filtered.
• The filtrate is acidified with Hcl.
• Sodium chloride is added to the acidified
filtrate to saturate it. NaCl causes
precipitation of enzyme.
• The precipitates are separated and dried.
• After drying it is converted into powder
form.
Description:
• Rennin is found in the form of powder, grain or
scales.
• The powder has yellowish white while grains and
scales have yellow color. it has a saline taste.
• It has a peculiar (special) odor which is not
unpleasant.
Standardization:
• After preparation the rennin is standardized
with reference rennin.
• The prepared rennin coagulates not less than
90% and not more than 110% of the
reference rennin.
• Rennin coagulates approximately 25000
times its own weight of fresh cow’s milk.
• If prepared rennin coagulates fresh cow’s
milk above this level, then lactose and sodium
chloride is mixed with it to bring its
strength to required strength.
• Rennin curdles of the milk i.e. It converts
the milk protein Casein into paracasein by
partial hydrolysis, Paracasein combine with
Ca+2 ion to form calcium Paracaseinate, which
is insoluble and is responsible for milk
curdling.
Uses:
• Rennin is used as digestant in pepsin and
rennin elixir.
• Rennin is used to coagulate the milk which
can easily be digested by convalescents.
• Rennin is used in the production and or
manufacturing of cheese.
2.Pepsin

• Pepsin is a substance containing a proteolytic

enzyme obtained from the glandular layer of
the fresh stomach of the hog,
Sus scrofa
Fam. Suidae
Preparation:
• Pepsin is prepared by digesting the minced
stomach linings with hydrochloric acid.
• This solution is clarified, partially
evaporated, dialyzed, concentrated, and
either poured on glass plates to dry, thus
forming scale pepsin,
• or carefully evaporated in a vacuum, forming
spongy pepsin.
Description:
• Pepsin occurs as transparent, or translucent
scales, as granular or spongy masses ranging
in color from light yellow to light brown, or
as fine white or cream-colored amorphous
powder.
• It is free from offensive odor and has a
slightly acid taste.
• Pepsin digests not less than 3000 and not
more than 3500 times its weight of
coagulated egg albumin.
• A pepsin of higher digestive power may be
reduced to the standard by admixture with a
pepsin of lower power or with lactose.
• Pepsin produced commercially, especially
spongy pepsin, often is 4 to 5 times as active
as that used medicinally.
Uses:
• Pepsin is administered to assist gastric
digestion.
• It is a proteolytic enzyme and should
preferably be given after meals.
• The usual dose is 500 mg.
• it is often combined with pancreatin in
product formulations.