This document discusses various proteins and enzymes used as pharmaceutical aids. It describes gelatin, casein, papain, bromelain, pepsin, urokinase, streptokinase, and serratiopeptidase. Gelatin is extracted from animal collagen and used in capsules and suppositories. Casein is the main protein in milk and used as an emulsifier. Papain from papaya, bromelain from pineapple, and pepsin from hog stomach are digestive enzymes. Urokinase from urine and streptokinase from bacteria are fibrinolytic enzymes. Serratiopeptidase from silk moth larvae is an anti-inflammatory enzyme. These naturally derived proteins and enzymes have various pharmaceutical

1. Proteins and enzymes as
Pharmaceutical aids
By Pradnya Salunkhe- Shinde

2. Proteins as pharmaceutical aids
• What are proteins?
• Gelatin
• Casein
• ENZYMES

3. Proteins
• Large molecules composed of one or more chains of
amino acids in a specific order determined by the
base sequence of nucleotides in the DNA coding for
the protein
• E.g. enzymes, hormones, and antibodies

4. Gelatin(gel foam)
• Biological source-
• It is preotein extracted by partial hydrolysis of animal
collagenous tissue like skins,tendons,ligaments and bones with
boiling water.
• Description_
• flakes, sheets, coarse or fine powder
• Charactristic odour
• Faint yellow to amber color

5. Preparation
• Bones defatted-dacalcified with organic solvent and
mineral acids resp
• Above material+water (at 85degree celcius)
• Collagen dissolves into gelatin
• Bleaching and cleaning
• Moulded it in treys
• dried

6. • Chemical constituents
• Amino acids-lysine
• Glutin protein
• Identification test
• Gelatin + soda lime----it evolves ammonia
• Gelatin+mercuric nitrate---white ppt---on warming it
give brick red color

7. Uses
• Hard and flexible capsule shape
• In different formulation preparation e.g.paste,
pessaries, suppossitories
• As a haemostatic
• Microencapsulation
• Culture media

8. Casein
• Biological source
• It is main phosphoprotein in milk and constitutes 3%
milk. It comprises about 80% total protein content of
milk.
• Two types of casein
• Acid casein
• Rannet casein

9. Description
• Color-white-slightly yellow
• Tasteless
• Odourless
• Amorphos solid
• Hygroscopic
• Stable when dried
• Deterorates rapidly
• Insoluble in water
• Soluble in dil.alkali

10. • Chemistry
• 15 amino acids
• 0.85%phosphorous
• 0.75%sulphur
• Mol wt.-75,000-3,70,000
• Isoelectric point-4.7
• Nitrogen content15-16%
• Standards
• LOD-6%
• Sulphated ash-NMT1.55
• Specific gravity1.25-1.31

11. Uses
• Diatery supplement
• Standardization of proteolytic enzymes
• Emulsifying agent
• Sizing of textile and paper
• As adhesive in paints

12. Enzymes
• Enzymes are proteins which acts as a biological catalyst
• Oxidoreductase: These enzymes catalyse oxidation and
reduction reactions.
• Transferase: These enzymes catalyse the transfer of
functional groups such as methyl or phosphate group.
• Hydrolases: These enzymes catalyse the hydrolysis
reactions.
• Ligases: These enzymes catalyse the joining of two
molecules by making a bond.
• Isomerases: These class of enzymes bring about the
isomerisation reaction
• Lyases: Lyases brings about lysis and cleavage of various
bonds.

13. Papain
• B.S.
• It is mixture of proteolytic enzyme derived from the
latex ofunripe fruits of tropical melon tree Carica
papaya belonging to family Caricaceae.

14. Method of preparation
• Latex of these fruit is collected
• Add potassium metabisulphate (5gm/kg)
• Extraneous matter cleared by passing through sievs
• Latex is dried in vaccum shelf at 55-60 ⁰c
• Spray dried (papain)

15. • Description
• Color-light brown to white color
• Typical odour and taste
• Max proteolytic activity at pH 5-6
• Soluble in H2O and glycerine
• Chemical constituets
• Papain+chymopappain

16. • Identification tests
It decolorizes aqu. potassium permanganate solution
• It causes curding of milk
• Uses
• Calrification of beverages
• Cheese manufacturing
• Degumming of silk fabric
• Leather industry for dehairing of skins
• Antiinflammatory agent

17. Bromelain
• B.S.It is mixture of proteolytic enzyme from stems and
ripen fruits of pineapple plant Ananas comosus, belonging
to family Bromeliaceae
• Activity-protein digesting and milk clotting enzyme
• Desription-
• odourless,
• buff colored with irritating taste

18. • Chemical constituents
• Peptidase, angin, cosmosain etc
• Vitamin
• Mono and disaccharides

19. Method of extraction
• Fruits-cut-weigh-macerate-juice
• Juice pressed and flitered(pH 6)
• Add ammonium sulphate until saturation to precipitate
the enzyme
• dissolve in sodium cyanide for purification
• Wash with ammonium sulphate then with acetone
• Thorough wash with acetone and ether
• Dried at low temperature.

20. • Solubility
• Soluble in water
• Insoluble in organic solvent like ether, chloroform,
alcohol
• Uses-
• Treatment of soft tissue inflammation and oedema due to
allergy and injury

21. Pepsin
• It is a substance containing proteolytic enzyme and
presentt in gastric juice of animals.
• Glandular layer (mucus membrane ) of fresh stomach
of hog Sus scrofa, vardomesticus, belonging to family
Suidae.

22. • Description
• Light buff/white color
• Acidic and saline taste with slightly meaty odour
• Soluble in water
• Insoluble in alcohol, ether and chloroform
• pH-1.8-max activity

23. • Preparation
• Stomach lining digested with HCl
• Clarification controlled evaporation, dialysis, conc.of
the digested solution
• Vaccum evaporation
• Spongy pepsin

24. • Therapeutic uses
• For proper digestion of food when patient lack its
secretion
• Patient .tiny bits and for further absorption as nutrients.
• Storage;-
• 2-8⁰c for one week at neutral pH under germ free
conditions.

25. Urokinase (uronase, winkinase)
• B.S.
• It is fibrinolytic enzyme found in the urine of cat, rat, cow man,
dog etc
• It is isolated from Baillus subtilis, Beta haemolytic streptococci
and urine sample of humans.
• Charactristcs-
• White to almost white color
• Fully soluble in water

26. Preparation/isolation
• Isolated by tissue culture technique(cell culture methods)
or recombinant DNA technology from kidney cells or
urinemainly obtained by human urine
• Uses
• Thromobolytic agent
• Treatment of pulmonary enzymes
• Treatment of acute ischemic stroke

27. Streptokinase
(estreptokinase/plasminokinase)
• It is purified bacterial proteolytic enzyme produced from
cultured filtrate of the strains of group C-Beta Haemolytic
S.Griseus
• Charactristic
• Sterile
• Friable
• Solid white
• Soluble in water

28. Preparation
• Isolated colonies of S.griseus singled out+25 ml sterilized
phosphate buffer pH7
• Place in orbital shaker at 120 rpm at 37⁰c on orbital
shaker for 24 hrs for innoculum preparation
• 5 ml innoculum added to 100 ml production media and
incubated at 37 ⁰c for 24 hrs on orbital shaker at 120 rpm
• Biomass harvested by filtration and centrifugation
• Supernatant purified (enzyme)

29. • Uses
• To dissolve blood clot
• For lysis of pulmonory embolism
• Storage
• Store and protected from light at -20⁰c

30. Serratiopeptidase(Serrapeptase)
• B.S.It is a proteolytic enzyme isolated from
nonpathogenic enterobacteria serratia E.15/serratiaa
marcescens
• This enzyme is produced in intestine of silk moth
larvae to breakdown cocci walls. It can thus be
obtained from the silk moth larvae.

31. Uses
• Reduces inflammatory oedema and swelling since
widely utilized as anti-inflammatory enzyme
• Superior alternative to NSAID drug

32. Thank you.