CLINICAL UTILITY OF ENZYMES

Dr Sukanya Shetty
Professor and Head
Department of biochemistry
Dates: 7th, 10th and 12th May 2022
 Instructions
• Studenta are required to prepare and come
 Specific learning objectives
At the end of the session, the learner should be able to
 Classify plasma enzymes based on functions

 Enumerate the difference between functional and non-functional

enzymes

 Classify enzymes based on clinical utility

 Define isoenzymes
 Specific learning objectives
At the end of the session, the learner should be able to

 Discuss the diagnostic importance of enzymes with examples

 Discuss the analytical importance of enzymes with examples

 Specific learning objectives

At the end of the session, the learner should be able to

 Discuss the role of enzymes as labels in diagnostic techniques such as
ELISA
 Discuss the role of enzymes molecular diagnostic techniques such as
PCR
 Enumerate the role of enzymes as tumor markers
 Discuss the therapeutic importance of enzymes with examples
 Explain the mechanisms of alterations of enzymes and isoenzymes in
disorder of heart , pancreas, bone and prostate
Classification of Plasma enzymes based on functions

There are two plasma enzymes

• Functional plasma enzymes

• Non- functional plasma enzymes

 Functional plasma enzymes
• Perform a physiologic function in blood.
• Their substrates are present in the circulation of normal individuals and
Examples: Lipoprotein lipase
pseudocholinesterase
proenzmes of blood coagulation.

• Generally they are synthesized in the liver but present in blood in

equivalent or higher concentration than in tissues.
 Non – functional plasma enzymes

• Perform no known physiologic function in blood.

• Their substrates are absent from plasma.

• Their levels are very low which is derived from normal destruction of
cells (tear and wear of the cells).

• Increased levels of these enzymes have diagnostic and prognostic

importance
Mechanisms responsible for abnormal levels of non- functional enzymes

1.Increase in serum levels

a) Necrosis of cells
b) Increased permeability of cell membrane
c) Increased production of these enzymes
d) Increased in tissue source
e) Decreased excretion
Decreased levels

1. decreased ceruloplasmin in wilsons disease.

2. decreased ALP in hypophosphatamia

3. decreased SGOT in lack of cofactors

4. decreased pseudocholisterase in hepatitis

 Examples for non-functional enzymes

• Lactate Dehydrogenase • GGT

• SGOT
• SGPT
• ALP
• CPK
• Amylase
• Lipase
• ACP
 Classification of enzymes based on clinical utility

• Diagnostic

• Analytical

• Therapeutic
 Isoenzymes

Different forms of the same enzymes synthesised by various tissues are

called isoenzymes or isozymes.
 Enzyme units

• Abbreviated as U/L

• One International unit is the amount of enzyme that will convert one
micromole of substrate per minute per liter of sample.
 Isoenzymes

• Isoenzymes (isozymes) are the physically distinct forms of the same

enzyme but catalyse the same chemical reactions.

• They differ from from each other structurally, electrophoretically and

immunologically.

• Isozymes are products of closely related genes.

 Examples for isoenzymes
Enzymes Isoenzymes Tissue
CPK CK-MM,CK-MB, CK-BB Muscle , myocardium, brain
LDH LDH1, LDH2, LDH3, LDH4 LDH5 Heart, RBC,Brain,Liver, muscle

ALP Biliary canaliculi, hepatic, placenta, Bone ,intestine

 Lactate Dehydrogenase (LDH)
Pyruvate Lactate
NADH + H+ NAD+

It has 5 isoenzymes
Isoenzyme subunits Electrophoretic Tissue Stability
mobility

LDH 1(30%) H4 fastest Heart Heat stable

muscle

LDH2 (35%) H3MI Faster RBC Heat stable

LDH3 (20%) H2M2 Fast Brain Heat labile

(partly)
LDH4 (10%) H1M3 Slow Liver Heat labile

LDH5 (5%) M4 slowest Skelatal Heat labile

muscle
 Lactate Dehydrogenase

Refernce range: 100 to 200 U/L

Increased in
• Hemolysis
• Myocardial infarction
• hemolytic anemias
• heptocellular injury
• muscular dystrophy
• carcinomas
• leukaemias
 Pattern in myocardial infarction

 Normally LDH 2 concentration is greater in blood than LDH1. but this is

reversed in myocardial infarction. This is called as flipped pattern.

 The serum level raises within 24hrs after infarction reaches a peak level
around 2-3 days and returns to normal in a week.
 Creatine phosphokinase

CPK
Creatine Creatine phosphate
ATP ADP
Isoenzyne Subunits Electrophoretic Tissue
mobility

CPK –1 BB Fast moving Brain

(1%)

CPK –2 (2 MB Intermidiate Myocardium

to 5%)

CPK --3 MM Slow moving Skeletal muscle

(90%)
 Creatine phosphokinase

Normal level : Males: Upto 190U/L

Females: Upto 170 U/L
Increased in
 Myocardial infarction
 hemolysis
 muscular dystrophy
 crush injury
 fracture
 Acute cerebrovascular accidents
CK and myocardial infarction
 The CK level start to rise within 3hrs of
infarction, reaches a peak within 24hrs
and returns to normal in a 3days time.

CK –mt :
 fourth isoenzyme
 located in the mitochondria
accounts for about 15% of total
activity.
 AST (Aspartate transaminase )
or
SGOT ( Serum Glutamate Oxaloacetate Transaminase )

Site - myocardial cells

liver cells

Reference range: 0 – 40 U/L

Increased levels
- myocardial infarction
- liver diseases
 Regan isoenzyme

• An isoenzyme closely resembling the placental form found in circulation

in conditions like carcinoma of lung, liver and intestine
 ALT (Alanine transaminase )
or
SGPT ( Serum Glutamate Pyruvate Transaminase )
Site - liver cells

Refernce range: 0 – 45 U/L

Increased levels
- liver diseases
Sudden fall in ALT in cases of hepatitis is a bad prognostic sign.
 Alkaline Phosphatase ( ALP )
Sites: liver, kidney, intestinal mucosa, bone and placenta
Reference range:
In children upper levels of normal level is seen because of increased
osteoblastic activity.
Moderate increase
 hepatitis
 hepatocellular carcinoma
Very high levels
• Cholestasis
• bone diseases
 Acid phosphatase
It hydrolyses the phosphoric acid esters at pH between 4 and 6.
Site– platelets, prostate, RBC and WBC.
Reference range: 1 to 5 KA units /100 ml
Isoenzyme
Prostate isoenzyme is tartrate labile

Clinical significance: total ACP level is increased in prostate cancer and

highly elevated in metastasis. In these conditions tartrate labile
isoenzyme is elevated.
This assay is useful in diagnosis and follow up treatment of prostate
cancers.
 Gamma Glutamyl Transpeptidase (GGT )

Site :liver, kidney, pancreas, intestinal cells

Reference range: 10- 30 U /l

Moderately elevated: infective hepatitis, obstructive jaundice

Highly elevated : alcoholism

 Amylase

 site : pancreas , salivary gland

 Reference range:

 Highly elevated in acute pancreatitis

 Moderately increased ion chronic pancreatitis, mumps etc

 Lipase

• Site – pancreatic secretion

• Normal level – 0.2 to 1.5 IU/L
• Significance - highly elevated in acute pancreatitis and persists for a
week.
 Ceruloplasmin ( Ferroxidase )

• Reference range:25 to 50 mg / dL

• Increased in inflammatory conditions

• Decreased in Wilsons disease

 Enzymes and diseases
Organs Routine Additional in
investigations special cases
Liver diseases SGOT, SGPT, ALP. GGT and 5’nucleotidase

Myocardial infarction CPK SGOT, & LDH.

Muscle diseases CPK & LDH aldolase

Bone diseases Alkaline phosphatase

Pancreatitis Amylase and Lipase

 Therapeutic role of enzymes
Streptokinase to lyse intravascular clot MI

Urokinase to lyse intravascular clot MI

Tissue plasminogen to lyse intravascular clot MI

activator
Pepsin, Trypsin Pancreatic
insufficiency
Asparaginase ant cancerous drug

α-1 antitrypsin emphysema

 Analytical uses
• Measurements of metabolites: Glucose oxidase, hexokinase, urease,
uricase

• Immunoassays: Alkaline phosphatase, horse radish peroxidase

• Recombinant DNA technology: Restriction endonuclease, tag

polymerase
 Analytical roles of enzymes

Glucose Estimation of glucose

oxidase/Hexokinase
Urease Estimation of urea

Cholesterol oxidase Estimation of cholesterol

Alkaline phosphatase ELISA

Restriction endonuclease RFLP, blotting techniques
Reverse transcriptase/Tag PCR
polymerase
Lipase TG