Pharmacogosy and phytochemistry-I(PCI syllabus Sem-IV)

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.