2. ENZYMES
• Enzymes are organic catalysts produced in the
body by living organisms. They perform many
complex chemical reactions that make up life
processes. Enzymes are lifeless and when
isolated, they still exert their characteristic
catalytic effect
• Enzymes are found in combination with
inorganic or organic substances that have an
important part in the catalytic action.
3. • Enzymes are obtained from plant and animal
cells and many have been purified. They are
used as therapeutic agents and as controlling
factors in certain chemical reactions in industry.
• Pepsin, pancreatin, and papain are used
therapeutically as digestants. Hyaluronidase
facilitates the diffusion of injected fluids.
Streptokinase and streptodornase dissolve
clotted blood.
4. Properties of Enzymes
1. Enzymes are sensitive to heat and are
denatured by excess heat or cold.
2. Enzymes are sensitive to pH, the rate at which
they can conduct reaction is dependent upon the
pH of where the reaction is taking place
3. Enzymes are reusable and some enzymes are
capable of catalyzing many hundreds or
thousands of reactions
5. 1. PEPSIN:
• Biological Source : It is the enzyme prepared
from the mucous membrane of fresh stomach
pig Sus scrofa Linn, belonging to family Suidae.
6. METHOD OF PREPARATION
• Mucous lining of stomach scrapped off
• Cut into small pieces
• Pulp placed in water
• Add HCL for digestion
• Kept at 37°C
7. • Autolysis takes place
• Clear liquid is obtained
• Contain peptones and pepsin
• Add NaCl
• NaCl precipitaes pepsin and peptone remain
as such
8. • Collection of ppts.
• Suspended in the water in dialyser
• Removel of salt by dialysis
• To the aqueous solution, add alcohol to bring ppt of
pepsin
• Collection of ppt
• Drying and packing
9. Description:
1. Colour: pale yellow
2. Odor: odourless or with very faint odour,
translucent grains
3. Taste: slightly bitter
4. It is best active at a temperature of 40°C with pH
2–4.
5. Pepsin is unstable above pH 6.
6. The enzyme gets denatured at a temperature of
70°C and in the presence of alcohol and sodium
chloride.
7. Pepsin can be stored for 1–2 years at 2–8°C.
10. Uses:
1. It is used in the deficiency of gastric secretion.
2. Pepsin is also used in the laboratory analysis of
various proteins; in the preparation of cheese,
and other protein-containing foods.
11. 2. UROKINASE
Synonym: Uroquinase.
Biological Source: Urokinase is serine protease
enzyme isolated from human urine and from
human kidney cells by tissue culture or by
recombinant DNA technology.
12. Preparation:
1. Urokinase is a fibrinolytic enzyme produced by
recombinant DNA using genetically manipulated
E. coli cells.
2. It is produced firstly as prourokinase and then
converted to active form by plasmin or kallikrein.
3. Urokinase used medicinally is also purified
directly from human urine.
13. 4. It binds to a range of adsorbents such as silica
gel or kaolin which can be use to initially
concentrate and purify the product.
5. It can be further purified by precipitation with
sodium chloride or ethanol or by chromatography.
6. Human urokinase needs sterile filtration,
aseptic filling and freeze drying.
14. Description:
1. Urokinase enzyme occurs in two different forms
as single and double polypeptide chain forms.
2. It has a half-life of 10–16 minutes after
intravenous administration.
3. It converts plasminogen to plasmin
15. Chemical Constituents:
1. Urokinase enzymes are serine proteases that
occur as a single low molecular weight (33 kDa)
and double, high molecular weight (54 kDa)
polypeptide chain forms. They differ in molecular
weight considerably.
Uses:
Urokinase is used in the treatment of pulmonary
embolism, coronary artery thrombosis and for
restoring the potency of intravenous catheters
16. 3.STREPTOKINASE
• Synonym: Estreptokinase, plasminokinase.
• Biological Source: It is a purified bacterial
protein produced from the strains of β-
haemolytic Streptococci.
17. Method of Preparation:
1. Streptokinase is a bacterial derived enzyme of
serine protease group.
2. Streptokinase is produced by fermentation
using streptococcal culture and is isolated from
the culture filtrate. It is produced in the form of a
lyophilized powder in sterile vials.
18. • Description: Streptokinase is a bacterial protein
with half-life of 23 minutes.
Chemical Constituents:
• Streptokinase is the purified bacterial protein
with about 484 amino-acid residues.
19. Uses:
1. Streptokinase is the first available agent for
dissolving blood clots.
2. Streptokinase and anistreptase are both used
in the treatment of pulmonary embolism,
venous, and arterial thrombosis and coronary
artery thrombosis.
20. 4.BROMELIN
Synonyms : Bromelin, bromelain.
Biological Source : Bromelin is a mixture of
proteolytic enzymes isolated from the juice of
Ananas comosus (pineapple), belonging to
family Bromeliaceae.
Geographical Source: It is grown in almost all
parts of the world including India, China,
Thailand, United States, Brazil, Philippines,
Mexico, Hawaii, and Taiwan.
21.
22. • Cultivation, Collection, and Preparation
• 1. Bromelin is found in pineapple fruit juice and stem.
• 2. It is propagated through suckers, slips, and crowns.
• 3. Plantation month is august.
• 4. Flowering month – February-march
• 5. fruit ripens during July–October
• 6. When fruits turns orange then they are cut off.
• 7. The enzyme bromelin does not disappear as the fruit
ripens. The enzyme from fruit and stem are known as
fruit bromelin and stem bromelin, respectively.
• 8. It is isolated from pineapple juice by precipitation with
acetone and also with ammonium sulphide.
23. Description:
• 1. The optimum pH of bromelain is 5.0–8.0.
• 2. In solution pH below 3.0 and above 9.5
inactivates the enzyme.
• 3. The optimum temperature is between 50 and
60°C.
• 4. The moisture content should not exceed 6%.
It is obtained in light brown coloured powder.
24. Chemical Constituents:
1. It is a mixture of sulphur-containing protein-digesting
enzymes, called proteolytic enzymes or proteases.
2. It also contains several other substances in smaller quantities,
including peroxidase, acid phosphatase, protease inhibitors, and
calcium.
Uses:
1. Bromelain is an effective fibrinolytic agent; bromelain inhibits
platelet aggregation and have anti-inflammatory effect.
2. Enhance healing process of body
3. as a digestive enzyme
4. as an antihypertensive
5. It may even be useful in the treatment of AIDS to stop the
spread of HIV.
25. 5.SERRATIOPEPTIDASE
• Synonym: Serrapeptase, serratiopeptidase.
• Biological Source: Serratiopeptidase is a
proteolytic enzyme isolated from
nonpathogenic Enterobacteria Serratia E 15. It
is also produced by the larval form of the silk
moth.
26. Preparation
1. Serratiopeptidase is produced by fermentation
technology by using nonpathogenic enterobacteria species
such as Serratia E 15.
2. The larvae of silk moth produce this enzyme in their
intestine to break down cocoon walls. It can thus be
obtained from the silk moth larvae.
Description:
1. When consumed in unprotected tablet or capsule, it is
destroyed by acid in stomach. However enteric coated
tablets facilitate its absorption through intestine.
Chemical Constituents: Serratiopeptidase is a proteolytic
enzyme of protease type XXVI.
27. Uses:
1. As an anti-inflammatory enzyme.
2. It is also used as a fast wound healing agent.
3. It is traditionally used to treat rheumatoid
arthritis and osteoarthritis.
4. It has wide ranging applications in trauma
surgery, plastic surgery, respiratory medicine,
obstetric and gynaecology.
28. 6.PAPAIN
Synonyms : Papayotin, vegetable pepsin
Biological Source
• Papain is the dried and purified latex of the
green fruits and leaves of Carica papaya
belonging to family Caricaceae.
29. Description:
1. Colour- white/brown colored amorphous
powder
2. Typical odour and taste
3. Soluble in water and insoluble in alcohol
4. Maximum proteolytic activity at pH between 5
to 6.
30. Preparation
• 1. The latex is obtained by making two to four
longitudinal incisions, by stainless steel or wooden
sharp knives.
• 2. The exudate is collected in nonmetallic
containers.
• 3. The latex is pass through sieves to remove dirt
and unwanted material.
• 4. Then to the latex Potassium metabisulphite is
added.
• 5. Mixture is spread on the trays and dried in the
vaccum shelf dryer at 55 to 60°C for 4 to 5 hours.
• 6. The dried latex is called papain.
31. • Chemical Constituents :
Papain
Proteolytic enzyme
1. Peptase (clotting enzyme)
2. Mixture of papain and chymopapain
3. Renin (used in preparation of cheese)
4. Peptidase-I (covert protein into dipeptide and
polypeptide)
32. Uses:
• 1. In clarification of beverages
• 2. It is meat tenderizer. It breakdown protein and
making the meat easier to chew.
• 3. In manufacturing of cheese as a substitute of
renin.
• 4. Deconing of silk febric in textile industry.
• 5. As dehairing agent in leather industry.
• 6. Medicinally, anti-inflammatory agent
• 7. Used as an ingredient in cleansing solution for
soft contact lenses
35. GELATIN
Synonyms: Gelfoam; puragel; gelatinum.
Biological Source: Gelatin is a protein derivative
obtained by evaporating an aqueous extract made
from bones, skins, and tendons of various domestic
animals. Some important sources are: Ox, Bos
taurus, and Sheep, Ovis aries belonging to family
Bovidae
.
36. PreparationThe process of manufacture of gelatin vary from
factory to factory. However, the general outline of the process is
given below.
Raw material
Bones, skins, and tendons of Bovideans is collected and
subjected to liming operation
Liming Process
The raw material is first subjected to the treatment known as
‘liming’. In this process, the skins and tendons are steeped for
fifteen to twenty and sometimes for 40 days in a dilute milk of
lime. During this, fleshy matter gets dis-solved, chondroproteins
of connective tissues gets removed and fatty matter is
saponified. The animal skin is further thoroughly washed in
running water.
37. Defattying: In case of bones, the material is
properly ground and defatted in close iron
cylinders by treatment with organic solvents such
as benzene. The mineral and inorganic part of the
bone is removed by treatment with hydrochloric
acid.
38. Extraction
The treated material from bones, skins and
tendons is boiled with water in open pans. The
clear liquid runs of again and again and is
evaporated until it reaches to above 45 per cent
gelatin content.
39. Setting: The concentrated gelatin extract is
transferred to shallow metal trays or trays with glass
bottom. It is allowed to set as a semisolid jelly.
Drying: The jelly is transferred to trays with a
perforated wire netting bottom and passed through
series of drying compartments of 30–60°C increasing
each time with 10°C. About a month is taken for
complete drying.
Bleaching: In case of darker colour, finished product
is subjected to bleaching by sulphur dioxide.
Bleaching affords a light coloured gelatin.
40. Description:
1. Color- colorless or pale yellow
2. Odor: Characteristic
3. Taste: Slight and broth like
4. Solubility: insoluble in cold water, but swells
and softens when immersed. Soluble in hot
water. Insoluble in alcohol, chloroform and
ether.
41. Chemical Constituents:
1. Gelatin consists of the protein glutin which on
hydrolysis gives a mixture of amino acids.
2. The approximate amino-acid contents are:
glycine, alanine, valine, cystine and cysteine
3. the adhesive nature of gelatin is due to the
presence of glutin.
42. Uses
1. Gelatin is used to prepare pastilles, pastes,
suppositories, capsules, pill-coatings, gelatin
sponge; as suspending agent, tablet binder,
coating agent, as stabilizer, thickener and
texturizer in food;
2. for manufacturing rubber substitutes,
adhesives, cements, and printing inks, plastic
compounds, artificial silk, photographic plates
and films,
3. for inhibiting crystallization in bacteriology, for
preparing cultures and as a nutrient.
43. 2.CASEIN
• Biological Source: Casein is a proteolytic
enzyme obtained from the stomachs of calves.
It is extracted from the proteins of the milk; in
the milk, casein is structured in voluminous
globules. These globules are mainly
responsible for the white colour of the milk.
44. Characteristics
• 1. The isoelectric point of casein is 4.6.
• 2. The purified protein is water insoluble. While
it is also insoluble in neutral salt solutions, it is
readily dispersible in dilute alkalis and in salt
solutions such as sodium oxalate and sodium
acetate.
• 3. Casein does not coagulate on heating. It is
precipitated by acids and by a proteolytic
enzyme (rennet).
45. Chemical Constituents
• 1. Milk consists of 80% of milk proteins
(casein).
• 2. The major constituents of casein are alpha
(s1) and alpha (s2)-caseins, β-casein and kappa-
casein. These caseins are conjugated proteins
with phosphate group(s) which are esterified
into serine residues they have a low solubility
at pH 4.6.
46. Uses
• 1. It is used in the manufacture of binders,
adhesives, protective coatings, plastics (such as
for knife handles and knitting needles), fabrics,
food additives, and many other products.
• 2. It is commonly used by bodybuilders as a
slow-digesting source of amino acids.
