This document discusses therapeutic and diagnostic applications of enzymes, isoenzymes, and coenzymes. It provides examples of how enzymes are used therapeutically to aid digestion, act as anti-clotting agents, and treat various conditions. It also discusses how enzymes are used diagnostically to detect levels of substances like glucose, liver enzymes, and more. The document then explains isoenzymes and provides an example of lactate dehydrogenase isoenzymes. Finally, it discusses several important coenzymes like NAD, FAD, biotin, vitamin B12 and their roles in biochemical reactions and maintaining health.

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
• Uro kinase Lyse Intracellular Clot
• Plasminogen Plasmin /Clot lysis
• Streptokinase DNA ase applied 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: Asparate aminotransferase (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 : Enzymes occurring in different molecular forms which differ in their physiochemical
properties but catalyze the same reaction.
Physio-chemical properties of Isoenzymes
1. differential mobility on electrophoresis
2. differential mobility in column chromatography
3. differential kinetic properties
Km, V max, Optimum temperature, Optimum pH, Relative sensitivity to inhibitors and degree of
denaturation.

8. Iso enzymes of Lactate dehydrogenase :

10. CO-ENZYMES

11. Definition
• Coenzymes are small molecules. They cannot by themselves catalyze a reaction but they can help enzymes to
do so. In technical terms, coenzymes are organic nonprotein molecules that bind with the protein molecule
(apoenzyme) to form the active enzyme (holoenzyme).
• The main coenzyme are NAD/NADP, FMN, FAD, Coenzyme A( CoA), Thiamine pyrophosphate, Pyridoxal
phosphate (PAL).

12. Salient features of coenzyme:
• Coenzymes are heat stable.
• They are low-molecular weight substances.
• The coenzymes combine loosely with the enzyme molecules and so, the coenzyme can be
separated easily by dialysis.
• When the reaction is completed, the coenzyme is released from the apo-enzyme, and goes to some
other reaction site.

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):
• Nicotinamide adenine dinucleotide (NAD) is a coenzyme found in all living cells. NAD+ was first discovered
by the British biochemists Arthur Harden and William John Young in 1906. It is consists of two nucleotides
joined through their phosphate groups. Nicotinamide adenine dinucleotide exists in two forms:
• An oxidized form (NAD+)
• Reduced form (NADH)
• NAD is called a dinucleotide because it consists of two nucleotides joined through their phosphate groups.
One nucleotide contains an adenine nucleobase and the other nicotinamide.

19. Biological functions and Importance
• Drug design and drug development exploits NAD+
• It is being studied for its potential use in the therapy of neurodegenerative diseases such as
Alzheimer's and Parkinson disease.
• NAD+ is also a direct target of the drug isoniazid, which is used in the treatment of tuberculosis.
• It is also used for the development of new antibiotics.

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-hydrofolic acid 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. Vitamin B12
• Vitamin B12, vitamin B-12, also called cobalamin, is a water-soluble vitamin with a key role in the
normal functioning of the brain and nervous system, and for the formation of blood.
• It is one of the eight B vitamins.
• It is normally involved in the metabolism of every cell of the human body, especially affecting
DNA synthesis and regulation, but also fatty acid metabolism and amino acid metabolism.
• Only bacteria and archaea have the enzymes required for its synthesis, although many foods are a
natural source of B12 because of bacterial symbiosis.

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