Therapeutic and diagnostic applications of enzymes isozymes and coenzymes

Therapeutic and diagnostic applications of Therapeutic and diagnostic applications of enzyme, Isoenzyme, Coenzyme

1. Therapeutic and diagnostic applications of
enzyme,
Isoenzyme,
Coenzyme
Presented By Shubhrat Maheshwari

2. Therapeutic applications of enzyme
1. Enzymes are used for aiding digestion. Example: Amylases, Proteases, Lipase.
2. They are used as deworming agent E.g.: Papain.
3. They act as anti-clotting agents like fibrinolytic and thrombolytic. Ex. Urokinase.
4. They act to treat atherosclerosis like serratio-peptidase.
5. They are used to treat wounds and swelling. Example: trypsin. Chymotrypsin,
serratio-peptidase.
6. They are used to assist metabolism like hyaluronidase.
7. They are used as surface disinfectants. Example: Trypsin

3. •Enzyme Therapeutic use
•Asparginase Acute Lymphatic Leukemia (cells need Asparagine for its growth )
•Streptokinase Lyse intracellular clot
•Urokinase Lyse Intracellular Clot
•Plasminogen Plasmin /Clot lysis
•Streptokinase DNA aseapplied locally
•Hyaluronidase Enhance local anesthesia
•Pancreatic (Lipase &
•Trypsin ) Pancreatic insufficiency –oral administration
•Papain Anti-inflammatory
•Alpha Anti Trypsin Emphysema

4. Diagnostic application
1. They are also used in the diagnosis purpose. Example: Glucose
oxidase along with peroxidase to detect the
level of glucose.
2. Liver disease: SGPT (gama-glutamyl transpeptidase)
3. Heart attacks: Asparateaminotransferase (AST)
4. Myocardial infection: Creatine phosphokinase
5. Uric acid: Uricase

5. ENZYME FOR DIAGNOSTIC PURPOSE ESTIMATION OF
UREASE UREA
URICASE URIC ACID
GLUCOSE OXIDASE GLUCOSE
PERIOXIDASE GLUCOSE /CHOLESTEROL
HEXOKINASE GLUCOSE
CHOLESTEROL OXIDASE CHOLESTEROL
LIPASE TRIGLYCERIDE
HORSE RADDISH PERROXIDASE ELISA
ALKALINE PHOSPHATASE ELISA
RESTRICTION ENDONUCLEASE SOURTHERN BLOT
REVERSE TRANSCRIPTASE POLYMERASE CHAIN REACTION

6. ISOENZYMES

7. Definition:Enzymesoccurringindifferentmolecularformswhichdifferintheirphysiochemical
propertiesbutcatalyzethesamereaction.
Physio-chemicalpropertiesofIsoenzymes
1.differentialmobilityonelectrophoresis
2.differentialmobilityincolumnchromatography
3.differentialkineticproperties
Km,Vmax,Optimumtemperature,OptimumpH,Relativesensitivitytoinhibitorsanddegreeof
denaturation.

8. Iso enzymes of Lactate dehydrogenase :

10. CO-ENZYMES

11. Definition
•Coenzymesaresmallmolecules.Theycannotbythemselvescatalyzeareactionbuttheycanhelpenzymesto
doso.Intechnicalterms,coenzymesareorganicnonproteinmoleculesthatbindwiththeproteinmolecule
(apoenzyme)toformtheactiveenzyme(holoenzyme).
•ThemaincoenzymeareNAD/NADP,FMN,FAD,CoenzymeA(CoA),Thiaminepyrophosphate,Pyridoxal
phosphate(PAL).

12. Salient features of coenzyme:
•Coenzymesareheatstable.
•Theyarelow-molecularweightsubstances.
•Thecoenzymescombinelooselywiththeenzymemoleculesandso,thecoenzymecanbe
separatedeasilybydialysis.
•Whenthereactioniscompleted,thecoenzymeisreleasedfromtheapo-enzyme,andgoestosome
otherreactionsite.

13. Thiamine pyrophosphate
•Thiamine pyrophosphate, or thiamine diphosphate, or co-carboxylase is a thiamine derivative which is
produced by the enzyme thiamine di-phosphokinase.
•Thiamine pyrophosphate is a cofactor that is present in all living systems, in which it catalyzes several
biochemical reactions such as Pyruvate dehydrogenase complex, pyruvate decarboxylase in ethanol
fermentation, alpha-ketoglutarate dehydrogenase complex etc.

14. Biological Function of Thiamine pyrophosphate:
It maintains:
The normal function of the heart;
Normal carbohydrate and energy-yielding metabolism;
The normal function of the nervous system;
Normal neurological development and function;
Normal psychological functions.

15. Flavin mononucleotide (FMN) and Flavin adenine
dinucleotide (FAD)
•Flavin is the common name for a group of organic compounds based on pteridine,. The biochemical source is
the riboflavin.
•It is commonly know as Vitamin B2. The flavin often attached with an adenosine diphosphate to form flavin
adenine dinucleotide (FAD),and, in other circumstances, is found as flavin mononucleotide (FMN).

16. Biological Functions and Importance:
Normal energy-yielding metabolism;
Normal metabolism of iron in the body;
The maintenance of normal skin and mucous membranes;
The maintenance of normal red blood cells;
The maintenance of normal vision;
The maintenance of the normal function of the nervous system.

17. NAD (Nicotinamide adenine dinucleotide)/NADP (Nicotinamide
adenine dinucleotide phosphate):
•Nicotinamideadeninedinucleotide(NAD)isacoenzymefoundinalllivingcells.NAD+wasfirstdiscovered
bytheBritishbiochemistsArthurHardenandWilliamJohnYoungin1906.Itisconsistsoftwonucleotides
joinedthroughtheirphosphategroups.Nicotinamideadeninedinucleotideexistsintwoforms:
•Anoxidizedform(NAD+)
•Reducedform(NADH)
•NADiscalledadinucleotidebecauseitconsistsoftwonucleotidesjoinedthroughtheirphosphategroups.
Onenucleotidecontainsanadeninenucleobaseandtheothernicotinamide.

19. BiologicalfunctionsandImportance
•DrugdesignanddrugdevelopmentexploitsNAD+
•Itisbeingstudiedforitspotentialuseinthetherapyofneurodegenerativediseasessuchas
Alzheimer'sandParkinsondisease.
•NAD+isalsoadirecttargetofthedrugisoniazid,whichisusedinthetreatmentoftuberculosis.
•Itisalsousedforthedevelopmentofnewantibiotics.

20. TH4 or Tetrahydro folic acid
Tetrahydro folic acid, or tetrahydrofolate, is a folic acid derivative.
Biological Functions:
It is a co factor in many reactions-
In the metabolism of amino acids and nucleic acids. A shortage in THF can cause megaloblastic anemia.
Tetra-hydrofolicacid is involved in the conversion of form iminoglutamic acid to glutamic acid
Reduce the amount of histidine available for decarboxylation and protein synthesis
It can decrease the urinary histamine and form iminoglutamic acid
THF

21. Pyridoxal pyrophosphate
•Pyridoxal pyrophosphate (pyridoxal 5'-phosphate), is the active form of vitamin B6, is a coenzyme in a
variety of enzymatic reactions.
•Vitamin B6 is a water-soluble vitamin.
•There are three different natural forms of vitamin B6: pyridoxine, pyridoxamine, and pyridoxal. Humans
depend on external sources to cover their vitamin B6 requirements.

22. Health Functions:
•It convert food into glucose, which is used to produce energy.
•It make neuro-transmitters, produce hormones, red blood cells, and cells of the
immune system.
•Along with vitamin B12 and vitamin B9, it control blood levels of homocysteine,
an amino acid that may be associated with heart disease.

23. Biotin
•Biotin is a composed of a ureido ring fused with a tetrahydrothiophene ring.
•Biotin is a coenzyme for carboxylase enzymes, involved in the synthesis of fatty acids, isoleucine, valine, and
in gluconeogenesis.
•Biotin, also known as vitamin H or coenzyme R, is a water-soluble B-vitamin (vitamin B7).

24. Biological role of Biotin:
A sufficient intake of vitamin B7 (biotin) is important as it helps the body to:
Convert food into glucose, which is used to produce energy.
Produce fatty acids and amino acids (the building blocks of protein).
Activate protein/amino acid metabolism in the hair roots and fingernail cells.
Maintain normal macronutrient metabolism.
The maintenance of normal. skin and mucous membranes.
The maintenance of normal hair

25. VitaminB12
•VitaminB12,vitaminB-12,alsocalledcobalamin,isawater-solublevitaminwithakeyroleinthe
normalfunctioningofthebrainandnervoussystem,andfortheformationofblood.
•ItisoneoftheeightBvitamins.
•Itisnormallyinvolvedinthemetabolismofeverycellofthehumanbody,especiallyaffecting
DNAsynthesisandregulation,butalsofattyacidmetabolismandaminoacidmetabolism.
•Onlybacteriaandarchaeahavetheenzymesrequiredforitssynthesis,althoughmanyfoodsarea
naturalsourceofB12becauseofbacterialsymbiosis.

26. Vitamin B12

27. Biological Role of Vitamin B12:
It maintain healthy nerve cells
It produce nucleic acids (e.g., DNA), the body's genetic material
It regulate the formation of red blood cells together with vitamin B9 (folate),
Together with vitamin B6 and vitamin B9, it control blood levels of the amino acid homocysteine,
a potential marker for heart disease risk.

28. Thank you