APPLICATION OF

ENZYME
EXAMINATION

BAIQ NASHA ISLAELI

NIM . 012024153011
 ENZYME
Enzymes are compounds produced by living cells which are
composed of proteins

enzymes function as biocatalysts, namely

accelerating the rate of chemical reactions.

The nature of the enzyme is specific to the substrate

basic enzyme examination in enzyme reactions with substrates
health in the cell

The presence of enzymes in body fluids

indicates that something has happened to the
cell walls, so that enzyme molecules can
penetrate them.
PURPOSE OF ENZYME
EXAMINATION
Support the diagnosis of a disease related to enzyme
activity

Know the location of the abnormality

Know the etiology of the disease

Follow up for a treatment

 AST

Transaminase
ALT

Creatinin Kinase
GGT
Enzyme
Examination
Alkaline
Posfatase

Oksidoreduktase LDH
AMINOTRANSFERASE
(TRANSAMINASE)
Catalyzes the
reversible transfer of
an amino group from
an amino acid to an
alpha-keto acid

Usually done for Example: ALT

liver function tests (SGPT), AST
(LFT) (SGOT), GGT
SGOT (ASPARTATE
AMINOTRANSFERASE)
Mediates the transfer of amino acid groups between aspartate and glutamate
EC 2.6.1.1

GOT (= Glutamate Oxaloacetate Transaminase) dimer + 2 PLP

(= pyridoxal phosphate, green) + 2 maleic acid (l.blue),
Metode : Kinetik (IFCC)
Prinsip :
2-Oxaloglutarate + L-Aspartate GOT L-Glutamate + Oxaloacetate
Oxaloacetate + NADH + H+ MDH Malat + NAD+

*Read Absorbance at a wavelength of 340 nm

* Normal value: 10-40 IU / L
Mild increase (1-3 times the normal value): Pancreatitis, fatty liver, biliary cirrhosis
Moderate increase (3-5 times the normal value): bile duct obstruction, arrhythmia,
liver tumor (metastasis), dystrophia muscularis
Moderate-severe increase (5 or more times the normal value): acute hepatocellular
damage, myocardial infarction, acute pancreatitis
SGPT (ALANIN
AMINOTRANSFERASE)
This enzyme plays a role in the alanine cycle
ALT catalyzes the reversible transamination between alanine and alpha ketoglutarate
to form pyruvate and glutamate.
EC 2.6.1.2
Metode : Kinetik (IFCC)
Prinsip :
2-Oxaloglutarate + L-Alanine GPT L-Glutamate + Piruvat
Piruvat + NADH + H+ LDH Lactate + NAD+

*Read Aborbance at a wavelength of 340 nm

* L-Alanine functions as an amino acid which will be converted into Glutamate by the
GPT enzyme
* LDH is also an enzyme that catalyzes the reaction of the product (pyruvate) being
converted into lactate.
* The reduction of Pyruvate to Lactate requires NADH and H +. NADH will undergo
oxidation to NAD +.
* The amount of NADH that is oxidized to NAD + is proportional to the number of the
GPT enzyme
CLINICAL MEANING
*Normal values: 5-35 IU / L

Mild increase (1-3 times the normal value): cirrhosis, alcohol abuse, pancreatitis,
fatty liver
Moderate increase (5-10 times normal): chronic liver disease, hemolysis, long-term
alcohol consumption
Weight gain (> 20 times normal): acute viral hepatitis, liver necrosis
ALT is considered to be more specific than SGOT for parenchymal damage to liver
cells. In general, the ALT test score is higher than AST in acute liver parenchymal
damage while chronic in the reverse
When ALT levels are high, other causes may be identified by measuring other
enzymes.
Example: elevated ALT due to liver cell damage can be distinguished from bile duct
obstruction by measuring the ALP enzyme.
GAMMA GLUTAMIL
TRANSAMINASE
γ-Glutamyl transferase (GGT) catalyzes the transfer of γ-glutamyl groups of
peptides, such as glutathione, to other amino acids.
GGT is a membrane enzyme found in hepatocytes and
biliary epithelial cells.
GGT activity in the serum is a marker of hepatobiliary EC : 2.3.2.2
injury, especially cholestasis and biliary effect, although it
is not only expressed in liver but also in kidney, pancreas,
and bile ducts.
GGT has been widely used as an index of liver dysfunction
and marker of high alcohol intake.

Struktur GGT
 GAMMA-GT
Metode : Kinetik Colorimetric
Prinsip :
L-gamma glutamil – 3 carboxy-4-nitroanilide + Glycilglycine GGT L-gamma glutamil-glycilglycine +
5-amino-2-nitrobenzoat
*Read absorbance at a wavelength of 405 nm
* Normal value: 0-40 U / L
Increased GGT activity in serum is usually an indication of hepatobiliary toxicity, especially
cholestasis.
GGT is elevated through induction in conditions of cholestasis and may be used to confirm
ALP activity elevations that are associated with cholestasis.
Some drugs and steroids can induce GGT such as Fenitoin, Barbiturat
ALKALINE FOSFATASE (ALP)
EC : 3.1.3.1
Catalyzes the reaction of removing phosphate groups from
proteins and from other molecules.

ALP includes Hydrolase enzymes

ALP is an enzyme that is produced mainly by the liver

epithelium and osteoblasts (new bone-forming cells).

Osteoblastic and osteoclastic activities increase ALP levels in ALP adalah enzim homodimerik
serum, these levels increase when there is bone growth or bone 86kDa. Setiap monomer
turnover mengandung 5 residu sistein
Metode : Optimized Standard Methode (DGKC)
Prinsip :
P-Nitrophenylphospate + H2O ALP Phosphate + P-Nitropheno
Substrate Produk

*Read absorbance at a wavelength of 405 nm

* Normal Value:
Male: 80-306 U / L
Female: 64-306 U / L
* elevated ALP levels can occur in primary billiard cirrhosis, bile duct obstruction
Electrophoresis can be used to distinguish Hepatic ALP
or Bone ALP.

The ALP isozyme is used to differentiate liver and

bone disease

ALP1 represents liver disease and ALP2 represents

bone disease

Other tests to distinguish liver ALP from other

isoenzymes are: 5'NT, Leusine Amnopeptidase (LAP),
and GGT.
LAKTAT DEHIDROGENASE
(LDH)
Catalyzes the reversible change of lactate to pyruvate

Including the oxidoreductase enzyme group

Laboratory panels for examination of heart abnormalities

LDH terdiri atas 4
subunit
The highest concentrations are found in heart muscle cells,
skeletal muscle, liver, kidneys, brain and red blood cells
• Metode : Kinetic
• Prinsip :
Piruvat + NADH + H+ LDH Lactate + NAD+

LDH has several isoenzymes:

LDH1: increase in myocardial infarction Reference Range : 120 -240 U/L
LDH2: increase in myocardial infarction
LDH3
LDH4: increase in liver infarction
LDH5: increase in liver infarction

* LDH levels increase due to megaloblastic anemia, hemolysis, muscular

dystrophy or infectious mononucleus.
CREATININ KINASE (CK)
The physiological role of CK is to maintain a large amount of phosphorylated
creatine energy, which is used to restore the amount of ATP that has been used up
during muscle contraction.
Reaction:

CK consists of 2 sub units, namely B and M. Each sub unit has a molecular weight
of 43 kDa. The combination of these 2 subunits produces only 3 isoenzymes.
Creatinine kinase (CK) is an enzyme that plays a role in muscle
metabolism.

CK has three isoenzymes, namely CK-MM, CK-MB, and CK-

BB.
EC : 2.7.3.2

CK-MM: Skeletal Muscles

CK-BB: Brain
CK-MB: Heart

CK-MB examination is used for acute myocardial infarction (AMI) and

for skeletal muscle disease / damage

CK MM, DIMER
PRINSIP PEMERIKSAAN
Metode : Optimized Standard Method
Prinsip : CK diaktifkan oleh N. Acetylcystein dengan reaksi :

Nilai normal :
Wanita : <110 U/L
Pria : <130 U/L
TAKE HOME MESSAGE
Enzyme tests can be used as biomarkers of a disease.
Types of enzyme tests:
1. liver enzymes: ALT, AST, GGT
2.Alkaline Phosphatase (ALP)
3.Creatinine Kinase (CK)
4.Lactate Dehydrogenase (LDH)