Enzymes

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 Objectives of presentation
 What are Enzymes?
 Some general properties of enzymes.
 Working of enzymes
 Classification of enzymes
 Some enzymes from natural origin
 Action of enzymes from natural origin
 Commercial products available.

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Introduction
Enzymes are organic catalysts produced
by living organisms. A lot of chemical
reactions carried out in Living organisms
are only possible due the presence of
enzymes.

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Introduction – Nature:
 Enzymes are biomolecules that catalyze chemical reactions.
 The vast majority of enzymes are proteins. In fact, until 1985, it was
believed that all enzymes were proteins. We now know that some RNA
molecules may also have catalytic activities and must thus be considered
enzymes too.
 In enzymatic reactions, the molecules at the beginning of the process are
called substrates, and the enzyme converts them into different
molecules, called the products. Almost all processes in a biological cell
need enzymes to occur at significant rates.
 Like all catalysts, enzymes work by lowering the activation energy
(Ea) for a reaction, thus dramatically increasing the rate of the reaction.
Most enzyme reaction rates are millions of times faster than those of
comparable un-catalyzed reactions.
 As with all catalysts, enzymes are not consumed by the reactions they
catalyse.
 However, enzymes do differ from most other catalysts by being much
more specific. 4
 Enzymes are known to catalyze about 4,000 biochemical reactions.
Introduction – Activity:
 Enzyme activity can be affected by other molecules.
 Inhibitors are molecules that decrease enzyme activity.
 Enzymes often occur in combination with organic or inorganic
substances that have an important part in the catalytic action. If these are
non protein organic compounds, they are known as coenzymes. If they
are inorganic ions, they are referred to as activators.
 Coenzymes are integral components of a large number of enzyme
systems. Several Vitamins(riboflavin, nicotinic acid) are recognized as
having a coenzyme function.
 Many drugs and poisons are enzyme inhibitors.

 Activity is also affected by temperature, chemical environment (e.g.,

pH), and the concentration of substrate.

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 Working of Enzymes
By definition, enzymes are biocatalysts. So, like other catalysts , enzymes also work by
lowering the Activation energy.
Activation energy:
Activation energy may be defined as the minimum energy required to start a
chemical reaction. The activation energy of a reaction is usually denoted by Ea and
given in units of kilojoules per mole.
The need for activation energy acts as a barrier to the chemical reaction occurring
and/or to the speed at which it occurs.
Enzymes lower the barriers that normally prevent chemical reactions from occurring(or
slow them down) by decreasing the required activation energy. Thus, in the presence of
enzymes, reactions proceed and/or proceed at a faster rate.

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 Classification
Enzymes are presently classified according to their action by a complex system
established by the commission on enzymes of the International Union of Biochemistry.
Six major classes are recognized which are as follows.
1. Oxidoreductases:
Catalyze oxido-reductions between two substances.
2. Transferases:
Catalyze transfer of a group, other than hydrogen, between a pair of substrates.
3. Hydrolases:
Catalyze hydrolysis of ester, ether, peptide, glycosyl, acid-anhydride, C-C, C-
halide or P-N bonds.
4. Lyases:
Catalyze removal of groups from substrates by mechanisms other than
hydrolysis, leaving double bonds.
5. Isomerases:
Catalyze interconversions of optic, geometric, or positional isomers.
6. Ligases:
Catalyze linkage of two compounds coupled to the breaking of a pyrophosphate
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bond in ATP or in similar compound.
Specificity:

Enzymes are usually very specific as to which reactions they catalyze and
the substrates that are involved in these reactions.

Responsible factors for the specificity of enzyme are:

Complementary shape,
charge and hydrophilic/ hydrophobic characteristics of enzymes and
 substrates.

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Lock & Key Model:
Enzymes are very specific, and it was suggested by Emil Fischer in 1894
that this was because both the enzyme and the substrate possess specific
complementary geometric shapes that fit exactly into one another. This is
often referred to as “the lock and key” model.
However, while this model explains enzyme specificity, it fails to explain
the stabilization of the transition state that enzymes achieve. The “lock and
key” model has proven inaccurate.

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Lock & Key Model:

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Induced Fit Model:
In 1958, Daniel Koshland suggested a modification to the lock and key
model:
Since enzymes are rather flexible structures, the active site is continually
reshaped by interactions with the substrate as the substrate interacts with the
enzyme.
As a result, the substrate does not simply bind to a rigid active site; the
amino acid side chains which make up the active site are moulded into the
precise positions that enable the enzyme to perform its catalytic function.
In some cases, such as glycosidases, the substrate molecule also changes
shape slightly as it enters the active site. The active site continues to change
until the substrate is completely bound, at which point the final shape and
charge is determined.
The induced fit model is the most currently accepted enzyme-substrate-
coenzyme figure.

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Induced Fit Model:

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Enzymes
A. Plant origin Enzymes:
a) Papain
b) Bromelain
c) Malt Extract
B. Animal origin Enzymes:
a) Renin
b) Pepsin
c) Pancreatin
d) Pancrelipase

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PAPAIN – Introduction:

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PAPAIN – Introduction:
Papain is the dried and purified latex of the fruit of papaya (Carica papaya)
Family (Caricaceae).
Papaya tree is indigenous to tropical America and is cultivated in Sri
Lanka, Tanzania, Hawaii and Florida. It is 5 – 6 m in height and fruit grows
to about 30 cm in length and up to 5 Kg weight. Papain is referred to as
vegetable pepsin because it contains enzymes somewhat similar to pepsin.
However unlike pepsin it acts in acidic, alkaline or neutral media.

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PAPAIN – Constituents:
S. N. Enzyme Action

1- peptidase 1 capable of converting proteins into

dipeptides and polypeptides,

2- a rennin like that act on casein of milk,

coagulating
enzyme
3- an amylolytic
enzyme

4- a clotting enzyme similar to pectase

5- an enzyme having feeble activity on fats.

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PAPAIN – Uses:
1) Papain can digest about 35 times its own weight of lean meat. It is sold
as a component in powdered meat tenderizer.
2) The best grade of papain digests 300 times its own weight of egg
albumin.
3) Papain, made into a paste with water, is also a home remedy for bee,
wasps stings, mosquito bites, breaking down the protein toxins in the
venom.

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BROMELAIN – Introduction:

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BROMELAIN – Introduction:

Bromelain is protein digesting and milk clotting enzyme obtained from the
juice of pineapple plant (Ananas comosus, Family Bromeliaceae).
Bromelain can appear in the juice of the fruit but it can also occur in the
stem of the plant . It differs from papain because it is obtained from both
ripe and inripe fruit.

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BROMELAIN – Uses:

1) Along with papain, bromelain is one of the most popular

substances to use for meat tenderizing.
Today, about 90% of meat tenderizer is used in consumer households.
Bromelain is sold in a powdered form, which is combined with a marinade,
or directly sprinkled on the uncooked meat. The enzyme will penetrate the
meat, and by a process called forking, cause the meat to be tender and
palatable when cooked. If the enzyme is allowed to work for too long, the
meat may become too “mushy” for many consumers’ preferences. As the
enzymes are heat labile therefore cooked or canned pineapple does not have
a tenderizing effect.

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BROMELAIN – Uses:

3) It is an anti-inflammatory agent, and so can be used for sports injury,

trauma, arthritis, and other kinds of swelling.
4) Its main uses are treatment of athletic injuries, digestive problems,
sinusitis, and aiding healing after surgery.

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MALT EXTRACT– Introduction:

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MALT EXTRACT– Introduction:
Barley is the dried grain of one or more varieties of Hordeum vulgare
(family Gramineae). Barley is grown throughout the worls where climate
is favourable.

Malt or malted barley : is dried artificially germinated barley grain. To

prepare malt, heaps of barley grain are kept wet with water in a warm room
and allowed to germinate. The grain is then quickly dried. The enzyme
diastase in the moist warm grains converts the starch to maltose, thereby
stimulating the embryo to growth. The embryo is killed when the grain is
dried.

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MALT EXTRACT – Introduction:
Dried malt resembles barley, but is more crisp, has an agreeable odour and
is more sweet in taste. It contains 50 – 70 % of sugar – maltose, 2 – 15 % of
dextrins, 8% proteins, diastase and a peptase enzyme.

Malt extract: is the product obtained by extracting malt. The malt is

infused with water at 60ċ and the expressed liquid is concentrated at a
temperature not exceeding 60ċ, preferably under reduced pressure.
Malt extract may be mixed with 10%, by weight, of glycerine. It contains
dextrin, maltose, a small amount of glucose, and amylolytic enzymes. It can
convert not less than 5 times its weight of starch into water soluble sugars.

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MALT EXTRACT– Uses:
1) Malt extract is used as an easily digested nutritive and as an aid in
digesting starch. Usual dose is 15g. Many commercial extracts of malt
do not contain diastase, which is destroyed by heat during sterilization.
2) Diastase is yellowish white, amorphous powder obtained from an
infusion of malt. It can convert 50 times its weight of potato starch into
sugars.
3) Also used as bulk producing laxative.

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PEPSIN – Introduction:
Pepsin is a digestive protease containing a proteolytic
enzyme obtained from the glandular layer of the fresh
stomach of hog – (Sus scrofa),
Family (Suidae)

It is an enzyme that is released by the chief cells in the

stomach and that degrades food proteins into peptides.
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PEPSIN – Preparation:
 Pepsin is prepared by digesting the minced stomach
linings with HCl. This solution is clarified, partially
evaporated, dialyzed, concentrated and either poured
on glass plates to dry – forming scale pepsin, or
carefully evaporated in vacuum forming spongy
pepsin.
 Pepsin occurs as lustrous, transparent or translucent
scales, as granular or spongy masses ranging in colour
from light yellow to light brown, or as fine white or
cream coloured amorphous powder. It is free from
offensive odour and has slightly acidic or saline taste.
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PEPSIN – Activity:
 Pepsin digests not less than 3000 and not
more than 3500 times its weight of
coagulated egg albumin.
 Pepsin functions best in acidic
environments, particularly those with a pH
of 1.5 to 2 and denatures if the pH is more
than 5.0.

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PEPSIN – Uses:
 Pepsin is administered to assist gastric
digestion. It is proteolytic enzyme and
should preferably be given after meals
 Usual dose is 500 mg.

 It is often combined with pancreatin in

product formulations.

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PANCREATIN – Introduction:

 Pancreatin is a mixture of several

digestive enzymes produced by the
exocrine cells of the pancreas.These
enzymes are amylase, lipase and
protease.
 This mixture is used to treat conditions in
which pancreatic secretions are deficient.

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PANCREATIN – Preparation:
It is obtained from the pancreas of hog – (Sus scrofa) or of the ox –
(Bos taurus). The pancreas is a gland that lies directly inside the posterior
wall of the abdomen. The fresh glands are minced and extracted by methods
similar to those employed in the manufacture of pepsin.

Pancreatin is a cream coloured amorphous powder with a faint,

characteristic but not offensive odour. Its greatest activity is in neutral or
faintly alkaline solution

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PANCREATIN – Activity:
Pancreatin contains in each mg, not less than 25
USP unit of amylase activity, not less than 2
USP units of lipase activity, and not less than 25
USP units of protease activity.
1USP unit of amylase activity is contained in the
amount of pancreatin that digests 1mg of dry
USP potato starch reference standard,
 1 USP unit of protease activity digests one mg
of casein, all under specified conditions.
1 USP unit of lipase activity liberates 1 μ Eq of
acid per minute at a PH of 9 and 37 c° . 32
PANCREATIN – Uses:

• Pancreatin is a digestive aid and is also

used in the preparation of pre-digested
foods for invalids.
• Enteric coated granules of pancreatin have
been used to treat infants with celiac
disease and related pancreatic deficiencies.
• Usual dose is 325 mg to 1gm, as tablet,
capsule or granules.

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PANCREALIPASE – Introduction:
Pancrealipase is essentially a more concentrated form of pancreatin, it
contains in each mg not less than 24 USP units of lipase activity, 100 USP
units of amylase activity, and 100 USP units of protease activity. Thus lipase
activity is increased 12 fold, but the amylase and potease activity only 4
fold, as compared to pancreatin.

Uses
Employed as a digestive aid, pancrealipase increases the intestinal
absorption of fat, thus aiding in the control of steatorrhea. It is available in
the form of capsules, powder packets and tablets. The usual dose ranges
8000 – 24000 USP units of lipolytic activity prior to each meal or snack, to
be determined by practitioner according to the needs of the patient suffering
from pancreatic insufficiency.
.

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RENNIN – Introduction:
Rennin is the partially purified milk curdling enzyme obtained from the
glandular layer of the stomach of the calf (Bos taurus) ,Family (Bovidae)

Preparation:
Rennin may be prepared by macerating the minced glandular layer of the
digestive stomach of the calf in 0.5 % NaCl solution, filtering, acidifying
the filterate with HCl and saturating with NaCl. The enzyme is precipitated
by the NaCl, separated, dried and powdered.
Rennin occurs as a yellowish white powder or as yellow grains or scales. It
has a characteristic and slightly saline taste and a peculiar, not unpleasant
odour. It is usually standardized so that it coagulates approximately 25000
times its own weight of fresh cow’s milk. 35
RENIN – Introduction:
Rennin can be used to coagulate milk, thus
rendering it more digestible for convalescents.
It is also an ingredient in pepsin and rennin
elixir.
 Its principal use, however, is to coagulate milk
for the manufacturing of cheese.

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