Biological source method of preparation ,identification test uses of enzymes

Phytochemical and pharmacognosy of enzymes

1. VidyabhartiCollege of pharmacy
2019-2020
Seminar Topic: Biological sources ,preparation ,identification test and uses of the following
enzymes.
Presented by : Mr. SushilMarotiraoBhoyar
B.Pharm. 4
th
year ( Sem. 7 )
Guided by : Prof. A. A. Deshmukhsir
( 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.
Biologicalsource: It is an aminolyticenzymes /mixture of enzymes obtained from rat pancreas, commercial it
is prepared from fungi i.eAspergillusOryzaeorbacteriaBacillus 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 diastaseis placed in platinum quarts or porceliancrucible.
Inthat 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 carbeneousmatter 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 hclis added to reciduewhich 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 etractof salt & Filtering it From conjugated protein &
precipitating the diastageby 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 starchyfood.
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 metasulphateis added in proportion of 5gm per kg of latex extraneous matters are
cleaned by passing it through sieve and the latex is dried in vaccumat 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. •Identificationtest :
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 SusScrofaLinn.
Family : Suidae.
Description :
Colour: Yellow to light Brown or as fine white
Nature : AmourphousPowder
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.
Pptsby addition of alcohol collect it, dried it.

9. Identification test :
Based on the protetolyticaction on it.
Coagulate egg albumin digesgestedwith 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 thecheese
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, PeptidylPeptide Hydrolase
Biological Sourse: It is the protiolyticenzyme obtained from mamalianpancreas of ox BosTaurus
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 trypsinhas been expressed many systems , including E-coli, Saccharomycescerevisaeand
PichiaPastoris.
PurificatIonocoursby affinity chromatography on benzamidineor 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 developewithin 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.Trypsingive to people who lack enzymes needed for digestion.
2.Also given with combination with bromelainand rutinfor 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, perubalsam , castor oil.).

12. Pencreatin
Synonym: Diastase Vera, Pendriotanone, Zypnar.
Biological Source: It is extracted from pancreas of certain animals like hog Sus-Sacrofa, Var
Domesticousbelonging to or ox Boss Taurus.
Family: Sudaeand 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 celsiusfor 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 Flurosenceunder UV lamp.
Uses :
Pancreatinis 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 mkemore tender (Sensitive
and Tasty ) before cooking ).

14. Reference
Textbook of Pharmacognosyand Phytochemistrypharmacopoeia and related drugs of natural
sources by Edward Jenner, page no.471-478.
C . K . Kokatepractical Pharmacognosy, VallabhPrakashan, Delhi.

15. Thank you...