Phytochemical and pharmacognosy of enzymes

1. Vidyabharti College of pharmacy
2019-2020
Seminar Topic : Biological sources ,preparation ,identification test and uses of the following
enzymes.
Presented by : Mr. Sushil Marotirao Bhoyar
B.Pharm. 4th year ( Sem. 7 )
Guided by : Prof. A. A. Deshmukh sir
( M. Pharm.)

2. INDEX
1. DIASTAGE
2. PAPAIN
3. PEPSIN
4. TRYPSIN
5. PANCREATIN

3. Introduction
 It is a protein which act as a catalyst
 They play vital role in a function of a cell and activities of the organism.
 Enzymes shows max. Activities b/w 35 – 40 Celsius.
 Inactive at 0 degree Celsius and beyond 65 Celsius denatured.
 The enzyme are protein that range in molecular weight from about 13,000 to as much as840,000
Dalton.
 Minute quantities of enzymes are required to complete a chemical reaction.
 Definition : the definition of enzyme is a protein created by an organism that increases rate at which
chemical reaction occur.

4. Diastage
 Synonym : α-Amylase & Maxylase.
 Biological source : It is an aminolytic enzymes /mixture of enzymes obtained from rat pancreas, commercial it
is prepared from fungi i.e Aspergillus Oryzae or bacteria Bacillus sup.
 Family : Graminae
 Descriptions :
1. Colour : White / cream colour.
2. Odour : Faint characteristics
3. Taste : Tasteless
4. Solubility : sparingly soluble in water, insoluble in alcohol & Ether
IDENTIFICATION TEST
 1) Aresenic:
 0.25 gm diastase is placed in platinum quarts or porcelian crucible.
 In that 10 ml of mg nitrate in ethyl alcohol is added and ignite it
 Then reduce to ash by heating at 450-550 degree celcious.
 If carbeneous matter persist it is wetted with minute amount of nitric acid which is further treated and heat at
a 4.0 to 5.0 degree celcious.
 After cooling 3ml of hcl is added to recidue which is dissolve by heating in a water bath.
 When this test is carried out.

5. Method of Preparation:
 It is obtained by heating cold water etract of salt & Filtering it From conjugated protein &
precipitating the diastage by adding the large quantity of absolute alcohol to the filtrate.
 It is further purified by resolution of water & Re-Precipitation with alcohol.
 Drying it done at 40 to 50 °c
Uses:
1. It is use as digestant.
2. It is used in pre-digested of starchy food.
3. It is also use as conversion of starch to fermentable sugar.
4. It is used in treatment of acid reflux tissue.
5. Distage catalyses the hydrolysis of a-1-4 glycoside linkage of polysaccharides.

6. Papain
Synonyms : Vegetable pepsin
Biological source : It is mixture of pyrolytic enzymes derived from the latex of unripe of tropical melon tree,
carica papaya linn.
Family : caricaccae
Methods Of Preparation :
1. Varticle incision are made on the fruit and the fruit latex is Collected In aluminum toys.
2. To This latex potassium metasulphate is added in proportion of 5gm per kg of latex extraneous matters are
cleaned by passing it through sieve and the latex is dried in vaccum at a temp of 55-60 C to obtain papain.
Description :
Colour : light in brown or white coloured
Odour : Characteristic
Tastes : Characteristic
Best active : pH of 5-6
solubility : soluble in water and glycerine.

7. •Identification test :
 Based on the proteolytic action of it.
 Coagulated egg albumin digested with a pepsin sol. and residue albumen tested with the reference.
Uses:
 It is the clarification of beverages, as a meat tenderiser, in cheese manufacture as a substitute of
renin.
 Acts as anti-inflammatory agent, It relieves symptoms of episiotomies.
 It’s also used in various industries like the textile industry for the degumming of a silk fabrics and
leather industries for deharing of skin.

8. Pepsin
 Biological Source : It is the enzyme prepared from mucous membrane of the stomach of various
animals like Pig , Sheep, Clap. The commonly used pieces of the pig is Sus Scrofa Linn.
 Family : Suidae.
 Description :
Colour : Yellow to light Brown or as fine white
Nature : Amourphous Powder
Odour : Slightly acid or saline taste.
 Method of preparation:
 Mucous membrane Is scraped from stomach and place in acidified water at 37c for 2 hrs.
 Filtered and sodium or ammonium salt till it become half saturated.
 Ppts by addition of alcohol collect it, dried it.

9. Identification test :
 Based on the protetolytic action on it.
 Coagulate egg albumin digesgested with a pepsin solution and residue albumin tasted with the
refrence.
Uses:
 It is used in the deficiency of gastric secretion.
 Pepsin is also used in the laboratory analysis of various protein; in the preparation of the the cheese
and other protein containing foods.
 It is also used in leather industry to remove the hair and residual tissue from hide.
 It is also used in preparation F2 fragments from antibodies.

10. Trypsin
 Synonym : Protinase, Peptidyl Peptide Hydrolase
 Biological Sourse : It is the protiolytic enzyme obtained from mamalian pancreas of ox Bos Taurus
 Family : Bovidae
 Description :
colour : White to whitish yellow
solubility: In water and insoluble in alcohol
Taste: Acrid
 Preparation :
 It is readily available from many source.
 Commercial sources contaminated with other pancreatic enzyme.
 Recombinant trypsin has been expressed many systems , including E-coli, Saccharomyces cerevisae and
Pichia Pastoris.
 PurificatIon ocours by affinity chromatography on benzamidine or aprotinin.

11. Identification test:
 Dil. 1 ml of solution S (Dissolve 0.10 gm in carbon monoxide free water and dilute 10 ml with same
solvent.) to 100 ml with water.
1. A reddish-violate colour develope within 3 min.
2. Dil. 0.5 ml of solution S to 5ml of solution water R .
3. Add 0’1 ml of a 20 gm /I solution of tosyl- lysyl- chloromethane hydrochloride.
4. Adjust pH 7 , shake for 2 hrs. and dil. 50 ml with water .....As above ......but negative test is
shown.
 Uses:
1. Trypsin give to people who lack enzymes needed for digestion.
2. Also given with combination with bromelain and rutin for treatment of osteoathritis.
3. Apply directly on wounds and ulcer to remove dead tissue and improve healing.
4. Spray-on product that used for healing mouth ulcers
(trypsin, peru balsam , castor oil.).

12. Pencreatin
 Synonym: Diastase Vera, Pendriotanone , Zypnar .
 Biological Source: It is extracted from pancreas of certain animals like hog Sus-Sacrofa , Var
Domesticous belonging to or ox Boss Taurus.
 Family: Sudae and Bovidae.
 Description: Fine amorphous powder.
 Colour: White to cream coloured.
 Odour: Faint characteristics odour.
 Preparation:
1. Pancreatic alpha- amylase: It is prepared by fractional precipitation aqueous ext. From hog or
bovine pancreas.
2. The technical enzyme preparation was used principally for textile sizing, because of their low
stability.
3. Pancreatic Alpha - amylase is used to prepare Pancreatin.

13. Identification test:
 50 mg. Of sub. In test tube, add 2-3 mg of resorcinol and 1 ml of Sulphuric acid , shake , heat, at
130 degree celsius for 5 min. and cool,
 Dilute with water ton 5ml and add hydroxide sol. Drop-wise to make it alkaline sol. , Cool 7 dil. To
10 ml of a greenish blue Flurosence under UV lamp.
 Uses :
 Pancreatin is used to treat pancreatic exocrine insufficiency (PEI) attributed to cystic fibrosis,
chronic Pancreatisis.
 In nutritional formula like tablets , capsules , or powder.
 In preparation of leather hides and meat tenderizer (added to meat to mke more tender (Sensitive
and Tasty ) before cooking ).

14. Reference
 Textbook of Pharmacognosy and Phytochemistry pharmacopoeia and related drugs of natural
sources by Edward Jenner, page no.471-478.
 C . K . Kokate practical Pharmacognosy , Vallabh Prakashan , Delhi.

15. Thank you...