ENZYMES IN MEDICINE

Diagnostic indicators the activities of many enzymes are

routinely determined in plasma ( rarely in tissue biopsies) for
diagnostic purposes in diseases of the heart, liver, skeletal muscle,
pancreas and other tissues - enzyme diagnostics

Therapeutic agents several enzymes are used as drugs; new

approach - enzymotherapy

Diagnostic tools use as chemicals in clinical laboratory assays

ENZYMES IN CLINICAL DIAGNOSIS

secretory - produced by tissues (namely
Enzymes

liver), acting in plasma

prothrombin, plasminogen, cerruloplasmin, choline

esterase; lipoprotein lipase

intracellular function intracellulary, have no physiological use in

plasma
- membrane bound ALP, GMT
- cytosolic
ALT, AST, LD, MDH
- mitochondrial
AST, GMDH
- lysosomal ACP

- tissue specific glucose-6-phosphatase liver

amylase pancrease
LD1 heart

Healthy individuals - levels of intracellular enzymes fairly constant, low

the rate of enzyme release from damaged cells into plasma balanced by the rate
of removal of enzyme protein from plasma

Physiological enzyme levels reference values of the enzyme activities

(determined in clinical laboratory each lab
has its own reference values)

Elevated enzyme activity in the plasma reflect tissue damage accompanied

by increased release of intracellular enzyme
Skeletal muscle during exertion physiologically elevated levels of muscle
enzymes in plasma

Many diagnostically important enzymes = isoenzymes pattern of

isoenzymes in plasma (determined electroforetically)
a means of identifying the damaged tissue

ALTERATION OF ENZYME PLASMA LEVELS

Increased values increased cell membrane permeability
anoxia, disturbances of energy metabolism
cytosolic enzymes ALT, LD, CK
- cell necrosis membrane-bound enzymes ALP, GMT
mitochondrial enzymes AST, GMDH
- induction of the enzyme synthesis drugs ALP, GMT

Decreased values inhibition of the activity drugs

- inhibition of the synthesis cell damage, drugs

Enzyme

EC number

Reaction

Use

Asparaginase

3.5.1.1

L-Asparagine H2O L-aspartate + NH3

Leukaemia

Collagenase

3.4.24.3

Skin ulcers

Glutaminase

3.5.1.2

Collagen hydrolysis
L-Glutamine H2O L-glutamate + NH3
Hyaluronate hydrolysis

Heart attack

Hyaluronidasea 3.2.1.35

Leukaemia

Lysozyme

3.2.1.17

Bacterial cell wall hydrolysis

Antibiotic

Rhodanaseb

2.8.1.1

S2O32- + CN- SO32- + SCN-

Cyanide
poisoning

Ribonuclease

3.1.26.4

RNA hydrolysis

Antiviral

-Lactamase

3.5.2.6

Penicillin penicilloate

Penicillin
allergy

Plasminogen plasmin

Blood clots
Inflammation
Blood clots

Streptokinasec 3.4.22.10
Trypsin

3.4.21.4

Uricased

1.7.3.3

Protein hydrolysis
Urate + O2 allantoin

3.4.21.31

Plasminogen plasmin

Urokinasee

Gout

Oldname

Newname

Amylase

Amylase

Normalranges

Approximate

OldNew

conversion

(various)
(i.u./1.)

factor
20

Lipase

Lipase

50180
1,000

Glutamic
oxaloacetic
transaminase

Aspartate
transaminase

4,000020
400

200
0.5

1035517
Glutamic
pyruvictransami
nase

Alanine
transaminase

825413

05

Lactic
dehydrogenase

Lactate
dehydrogenase

12050060
250

05

a
hydroxybutyrate
dehydrogenase

2
hydroxybutyrat
e
dehydrogenase

10030050
150

Isocitric
Dehydrogenas

Isocitrate
dehydrogenase

Aldolase

Aldolase

05
31013
5
03
210157

07

Examples of enzymes commonly assayed for diagnostic purposes

Enzyme

Location

Cause of elevated plasma level

Acid phosphatase - ACP

Prostate

Prostatic cancer

Alkaline phosphatase ALP

Bone, liver

Rickets, hypoparathyroidism,
osteomalacia, obstructive
jaundice, cancer of bone/liver

Alanine aminotransferase ALT

Liver (muscle,
heart, kidney)

Hepatitis, jaundice, circulatory

faillure with liver congestion

Aspartate aminotransferase AST Heart, muscle,

Myocardial infarction, muscle

red cells, liver

damage, anemia, hepatitis,

circulatory faillure with liver
congestion

Amylase - AM

Pancres

Acute pancreatitis, peptic ulcer

-Glutamyl transferase GMT

Liver, kidney,
pancreas

Hepatitis, alcoholic liver

damage, cholestasis

Examples of isoenzymes commonly assayed for diagnostic purposes

Enzyme
Creatine kinase CK
CK-MB
CK-MM
Lactate dehydrogenase LD
LD1 > LD2

LD2, LD3
LD5

Location

Cause of elevated plasma level

Heart
Skeletal muscle

Myocardial infarction
Muscular dystrophy

Heart, kidney,
blood cells

Myocardial infarction, kidney

disease, megaloblastic anemia,

leukemia
Leukemia
Liver, muscle
Liver disease, muscle damage

ENZYMES IN THERAPY

Substitution of missing production of digestive enzymes digestive

enzymes pepsin trypsin

Removal of deposits of death tissue or fibrin (e.g. in lungs, eyes),

treatment of skin defects proteinases, nucleases, collagenase

Acceleration of fibrinolysis in lungs embolization (activation of plasmin

and plasminogen) streptokinase, urokinase

ENZYMOTHERAPY
Orally administered enzymes treatment of a variety disorders
- digestive, gastrointestinal, pancreatic
- inflammatory diseases, edema
- immune and autoimmune diseases
(arthritis, multiple sclerosis)
- viral diseases (herpes, AIDS)
- cancer
Mixtures of enzymes of plant and/or animal origin - proteinases, amylase,
lipase - administered as acidoresistent tablets

Pancreatin trypsin, chymotrypsin, lipase, amylase

Wobenzym pancreatic and plant proteolytic enzymes trypsin,
chymotrypsin, papain (Carica papaya), bromelain (ananas) =
combination of enzymes with different specificity, pH optimum,
stability, interaction with inhibitors and antiproteinases
multiple action

 Mechanism of resorption (transport of large macromolecules across

the intestinal barrier) paracellular transport, receptor mediated
endocytosis and transcytosis
Mechanism of action interaction with plasma antiproteinases
1-antitrypsin, 2-macroglobulin complexes

direct proteolytic action, degradation of adhesive

molecules, secretion of cytokins (tranforming growth factor
TGF-), modulation of receptor function

not fully clarified

ENZYMES - USE IN LABORATORY ASSAYS

Enzymes isolated from different sources - used for determination of various
substances in the blood, plasma/serum and urine enzyme methods
much more specific than chemical methods, the presence of relative
substances with similar chemical properties does not hinder
Components of commercial kits or diagnostic strips
- determination of glucose - glucose oxidase, peroxidase
cholesterol - cholesterol esterase, cholesterol oxidase
peroxidase,
urea urease, .
in blood, plasma, serum
- proof of glucose (glucose oxidase), ..
in blood or urine (strips)
Markes in the immunochemical analysis
- ELISA (=enzyme-linked immunoadsorbent assay) peroxidase, alkaline
phosphatase