Hybridoma Technology ( Production , Purification , and Application )
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Proteolytic enzyme
1. PHARMACOGNOSY OF PROTEOLYTIC ENZYMES
Academica In-Charge, HOD,
PritamJuvatkar
Mobile :
Email : pritamjuvatkar@gmail.Com
9987779536
Department of Pharmacognosy and Phytochemistr
Konkan Gyanpeeth Rahul Dharkar College of
Pharmacy and Research Institute, karjat
3. Biological source:
• Serratiopeptidase is a protiolytic enzyme excreted by nonpathogenic Enterobacteria serratia
isolated from intestine of silkworms Bombyx mori L.
• It has a distinctive ability to digest the dead tissue of the Cocoon of the silkworm.
Macroscopical characters:
• Colour: A grayish white to pale brown colour powder with
• Odour: characteristic odour.
9987779536
Pritam Vijay juvatkar
PHARMACOGNOSY OF PROTEOLYTIC ENZYME
Serratiopeptidase
4. • In human body it breaks down protein deposits such as fibrin.
• This is used as a natural alternative to steroids and NSAIDs without serious side effects.
• The enzyme causes proteolysis of all non-vital tissues including blood clots, cysts, tissue
plaques and cellular debris and reduces the inflammatory response.
9987779536
Pritam Vijay juvatkar
PHARMACOGNOSY OF PROTEOLYTIC ENZYME
Serratiopeptidase
5. 9987779536
Pritam Vijay juvatkar
Serratiopeptidase binds to α2 – macroglobulin in the blood in 1:1 ratio.
It is slowly transferred to the exudates at the site of infection and inflammation.
Mechanism of action is by hydrolysis of histamin, bradykinin and serotonin.
By hydrolyzing bradykinin, histamine and serotonin it indirectly reduces dilatation of blood capillaries and controls
permeability.
Serratiopeptidase blocks plasmin inhibitors thus helping the fibrinolytic activity of plasmin.
Degradation of extra-fibrin to small fragment prevent clogging of microcapillaries, helps clearance of exudates, reduces
swelling and improves microcirculation & expectoration of sputum etc
Serratiopeptidase MOA
6. Uses
• Pain, inflammation due to
• Arthritis,
• Trauma,
• Surgery,
• Sinusitis,
• Bronchitis,
• Carpal tunnel and painful swelling of the breasts
• The recommended dosage for Serratiopeptidase is 10 mg to 30 mg a day.
• Serratiopeptidase is well known for its ability to reduce pain by blocking the release of paininducing molecules
from inflamed tissues
9987779536
Pritam Vijay juvatkar
PHARMACOGNOSY OF PROTEOLYTIC ENZYME
7. Biological source:
• Urokinase was first discovered and isolated from human urine [3].
• The urokinase activity in urine is elaborated from the cells lining the kidney tubules [2].
• The urokinase is secreted by vascular endothelial, smooth muscle cells (SMC), epithelial cells, fibroblasts,
monocytes/macrophages, and also by cells of malignant tumors of different origin [4–6].
• Cultured cells have been reported to secrete larger quantities of urokinase (50–100 ng/ml) as compared to that present in urine
(10–15 ng/ml) [7]. Human plasma also contains at least 10 g/L of urokinase-reactive material [8].
• Macroscopical characters:
• Colour: A white powder
• Odour: Characteristic odour.
9987779536
Pritam Vijay juvatkar
PHARMACOGNOSY OF PROTEOLYTIC ENZYME
Urokinase
8. 9987779536
Pritam Vijay juvatkar
Urokinase structure is subdivided into three domains: the N-terminal epidermal “growth factor-like domain” (GFD) is responsible for
the interaction of urokinase with uPAR/CD87 receptor
The C-terminal protease domain of urokinase includes the active site of the enzyme represented by amino acid triad His204, Asp255,
and Ser356, specific for serine proteases;
The kringle domain of urokinase interacts with plasminogen activator inhibitor 1 (PAI-1)
Each domain of the protein has a rigid structure supported by internal disulfide bonds.
Structure and function of urokinase
Site-specific mutagenesis studies have shown that only the disulphide bridges in the catalytic domain are essential for maintaining
amidolytic and Fibrinolytic activities of the plasminogen activate
9. Uses
• It is used to dissolve fibrin or blood clots in anterior chamber of eye and in acute massive pulmonary
emboli.
• As it is derived from human souce it is less antigenic than enzymes with similar actions like
streptokinase.
9987779536
Pritam Vijay juvatkar
PHARMACOGNOSY OF PROTEOLYTIC ENZYME
10. Biological source:
• It is an enzyme obtained from culture filtrates of B-hemolytic Streptococci group C.
• Macroscopical characters:
• Colour: Sterile, friable solid or white powder
• Solubility: Soluble in water with maximum activity at pH 7
• Stability: The solution at higher concentrations is stable for 6 hours at 4°C, otherwise dilute solutions are unstable.
9987779536
Pritam Vijay juvatkar
PHARMACOGNOSY OF PROTEOLYTIC ENZYME
Streptokinase
11. Uses
• It is used in the treatment of thromboembolic disorders for the lysis of pulmonary emboli, arterial thrombus, deep vein
thrombus and acute coronary artery thrombosis.
• The activity of this enzyme is due to activation of plasminogen to a proteolytic enzyme namely plasmin which degrades
fibrin clots, fibrinogen and other plasma proteins.
Dosage and Administration
• The most widely used and currently recommended dosage of intravenously administered streptokinase in elderly patients
with acute myocardial infarction is 1.5 million units infused over 30 to 60 minutes.
• Therapy should be commenced as soon as possible after the onset of symptoms, although a significant reduction in
mortality has been demonstrated with streptokinase administration up to 12 hours after symptom onset
9987779536
Pritam Vijay juvatkar
PHARMACOGNOSY OF PROTEOLYTIC ENZYME
12. Biological source:
• It is substance containing proteolytic enzyme and is present in the gastric juice of animals.
• It is obtained from the glandular layer (mucous membranes) of fresh stomach of hog, Sus scrofa var domesticus
Family : Suidae.
• Macroscopical characters:
• Colour: Light buff or white colored amorphous powder
• Shape: As translucent scales
• Odour: Slightly meaty odour
• Taste: Little acidic or saline taste
• Stability: Soluble in water, but insoluble in alcohol, ether and chloroform.
• Stability: If pepsin is heated with alkali or pancreatic enzymes, its biological activity is lost. It shows maximum activity at pH 1.8.
PHARMACOGNOSY OF PROTEOLYTIC ENZYME
Pepsin
13. Uses
• Pepsin has the capacity to digest 2500 its weight of coagulated egg albumin.
• It is also available in other forms which may digest even up to 10,000 times their weight of coagulated egg
albumin.
• For preparation, the minced stomach linings are digested with hydrochloric acid followed by clarification,
controlled evaporation, dialysis and concentration of the digested solution.
• When processed, solution is subjected carefully to vacuum evaporation, spongy Pepsin is obtained. Pepsin
degrades proteins into peptones and proteases.
9987779536
Pritam Vijay juvatkar
PHARMACOGNOSY OF PROTEOLYTIC ENZYME