SWU-PHINMA

COLLEGE OF
PHARMACY
ENZYMES TERMS DEFINITION
simple enzyme is an enzyme composed only of protein (amino acids) not bound to any nonproteins
Greek word en “in” and zyme “yeast”
Enzymes are biological catalyst that speed up the rate of the biochemical reaction. conjugated enzyme is an enzyme that has a nonprotein part in addition to a protein part
Most of the enzymes are three dimensional globular proteins (tertiary and quaternary structure)
apoenzyme is the protein part of a conjugated enzyme or holoenzyme
Note: With the exception of a few catalytic RNA molecules, or ribozymes which made of ribonucleic acids and that catalyze
enzyme lacking an essential cofactor
cellular reactions involving nucleic acids.
cofactor is the nonprotein part of a conjugated enzyme or holoenzyme
serve functions similar to those of prosthetic groups but bind in a transient, dissociable manner either to the
BIOCHEMICAL IMPOTRTANCE of Enzymes enzyme or to a substrate such as ATP.
Biological catalyst
prosthetic group Tightly bound cofactor to the apoenzyme
- They increase the rate of chemical reactions taking place within living cells without changing themselves.
- As catalyst, enzymes are not consumed during the reaction but merely help the reaction occur more rapidly. holoenzyme is the biochemically active conjugated enzyme produced from an apoenzyme and a cofactor
Specificity Intact and functional enzyme containing all cofactors/ coenzyme
- A given enzyme is very selective
- Both in the substances with which it interacts and in the reaction that it catalyzes Coenzyme or is a small organic molecule that serves as a cofactor in a conjugated enzyme
cosubstrate
Regulate metabolism
- play key role in the degradation and synthesis of nutrients: ex. digestion activator the inorganic cofactor
- presence and maintenance of a complete and balanced set of enzymes is essential for the breakdown of
nutrients to supply energy or to harness energy to power cell motility and activity Substrate (S) is the reactant in an enzyme-catalyzed reaction
biomolecule that enzymes react with
Synthesis of biomolecules
- the assembly of those building blocks into proteins, DNA, membranes, cells, and tissues Product (P) the biomolecules formed by enzyme mediated reactions

metal-activated enzymes Enzymes that require a metal ion cofactor

Enzyme and substrate interaction
metalloenzymes. enzymes that contain tightly bound metal ions

ENZYME

PROSTHETIC GROUP CONJUGATED ENZYME SIMPLE ENZYME

HOLOENZYME COFACTOR APOENZYME

 Cofactor – is the nonprotein part of the
conjugated enzyme. It is generally either a small organic
molecule or an inorganic ion (usually a metal ion) ACTIVATOR COENZYME
 Coenzyme are organic non-protein compound
that binds with an enzyme to catalyze a reaction. Just like
enzymes can be reused and recycled without changing
reaction rate or effectiveness. METAL ION
 Prosthetic Groups are distinguished by their
tight, stable incorporation into a protein’s structure by
covalent or noncovalent forces METAL-ACTIVATED ENZYME

METALLOENZYME
 COENZYMES NOMENCLATURE and CLASSIFICATION OF ENZYME
Vitamins Coenzyme Group transferred A. Based upon the substance they act upon (substrate)
 the suffix “ase” is added to the name of the substrate Except for:
Vit. B1 Thiamin pyrophosphate (TPP) aldehydes 1. amylase acts on starch Pepsin – in gastric juice (stomach)
Thiamin 2. maltase acts on maltose Trypsin – in pancreatic juice (small intestine)
3. cellulase act on cellulase Ptyalin – in saliva
Vit. B2 flavin mononucleotide (FMN) hydrogen atoms 4. lipase acts on fats Rennin – the milk curding enzyme
Riboflavin flavin adenine dinucleotide (FAD)
B. Based upon the reaction enhanced
Vit. B3 nicotinamide adenine dinucleotide (NAD1) hydrogen atoms  the suffix “ase” is added to the type of chemical reaction activated
Nicotinic acid nicotinamide adenine dinucleotide phosphate (NADP1) 1. Oxidase – oxidation reaction
2. decarboxylase - decarboxylation
Vit. B5 coenzyme A (CoA) acyl groups 3. transaminase - transamination
Pantothenic acid 4. hydrolase - hydrolysis
except: urease – hydrolysis of urea
Vit. B6 pyridoxal-5-phosphate (PLP) amino groups
lactase – hydrolysis of lactose
Pyridoxine pyridoxine-5’-phophate (PNP)
sucrase – hydrolysis of sucrose
pyridoxamine-5’-phosphate (PMP)
cellulase – hydrolysis of cellulose
Vit. B7 biotin carbon dioxide (carboxyl group) 5. hydrase or dehydrase – reversible addition or removal of water
Biotin
C. Based upon the reaction they catalyze
Vit. B9 tetrahydrofolate (THF) one-carbon groups other 1. OXIDOREDUCTASE
Folic acid than CO2 • Oxidation-Reduction reaction
Example: Lactate dehydrogenase
Vit. B12 Methylcobalamin (Cobalt as the prosthetic group metal ion) methyl groups, hydrogen Phosphate dehydrogenase
Cyanocobalamin atoms

METALLOENZYME
Metal ion Enzyme Function

Cu2+, Zn2+ Superoxide dismutase Destroy superoxide anion

“browning reaction” caused by phenolase (or polyphenoloxidase), a conjugated enzyme in which copper is present “
Zn2+ Alcohol dehydrogenase They oxidize a range of aliphatic and aromatic alcohols to their corresponding aldehydes and
ketones using NAD+ as a coenzyme. 2. TRANSFERASES
 is an enzyme that catalyzes the transfer of a functional group other than hydrogen (ex. methyl, acyl, amino or
DNA polymerase Synthesize DNA phosphate groups) from one molecule to another.
Carboxypeptidase A is a protease enzyme that hydrolyzes (cleaves) a peptide bond at the carboxy-terminal (C-
terminal) end of a protein or peptide Examples: transaminases and kinases

Mn2+ Pyruvate carboxylase Synthesize oxaloacetate

Arginase Synthesize Urea

Fe Hemoglobins transports oxygen from the lungs to the capillaries of the tissue.

Cytochromes carry electrons between two segments of the electron-transport chain.

Cytochrome P450 enzymes play a role in the synthesis of many molecules including steroid
hormones, certain fats (cholesterol and other fatty acids), and acids used to digest fats (bile
acids). Additional cytochrome P450 enzymes metabolize external substances, such as
medications that are ingested, and internal substances, such as toxins that are formed within
cells.

Cu2+ Cytochrome Oxidase hydrolyzes peptide bonds of C-terminal residues with aromatic or aliphatic side-chains
3. HYDROLASE Main Classes and Subclasses of Enzymes
 an enzyme that catalyzes a hydrolysis reaction in which the addition of a water molecule to a bond causes the
bond to break
 Hydrolysis reactions are central to the process of digestion. Main Class Selected subclasses Type of Reaction Catalyzed
Examples:
 Carbohydrases - breaking of glycosidic bonds in oligo- and polysaccharides OXIDOREDUCTASES oxidases oxidation of a substrate
 Proteases- breaking of peptide linkages in proteins reductases reduction of a substrate
 lipases - breaking of ester linkages in triacylglycerols dehydrogenases introduction of double bond (oxidation) by formal removal of two H atoms
from substrate, the H being accepted by a coenzyme

TRANSFERASES Transaminases transfer of an amino group between substrates

Kinases transfer of a phosphate group between substrates

HYDROLASES Lipases hydrolysis of ester linkages in lipids

Proteases hydrolysis of amide linkages in proteins
Nucleases hydrolysis of sugar–phosphate ester bonds in nucleic acids
Carbohydrases hydrolysis of glycosidic bonds in carbohydrates
phosphatases hydrolysis of phosphate–ester bonds

LYASES dehydratases removal of H2O from a substrate

decarboxylases removal of CO2 from a substrate
4. LYASE
deaminases removal of NH3 from a substrate
 is an enzyme that catalyzes the addition of a group to a double bond or the removal of a group to form a double hydratases addition of H2O to a substrate
bond in a manner that does not involve hydrolysis or oxidation.
Example: ISOMERASES Racemases conversion of D isomer to L isomer,or vice versa
 Dehydratase effects the removal Mutases transfer of a functional group from one position to another in the same
of the components of water from molecule
a double bond
 Hydratase effects the addition of LIGASES Synthetases formation of new bond between two substrates, with participation of ATP
the components of water to a Carboxylases formation of new bond between a substrate and CO2, with participation of
double bond ATP

5. ISOMERASE
 an enzyme that catalyzes the
OVERVIEW: MECHANISM OF ENZYME ACTION
isomerization (rearrangement of atoms or
interconversion of optical, geometric, or
positional isomers) of a substrate in a reaction,
converting it into a molecule isomeric with
itself. There is only one reactant and one
product in reactions where isomerases are
operative.

6. LIGASE or SYNTHETASE
• an enzyme that catalyzes the
bonding together of two Enzyme Active Site
molecules into one with the - the relatively small part of an enzyme’s structure that is
participation of ATP. involved in catalysis. Usually a “crevicelike” location the
• ATP involvement is required enzyme.
because such reactions are
generally energetically
unfavorable and they require the Enzyme-Substrate Complex
simultaneous input of energy - the intermediate reaction species that is formed when the
obtained by a hydrolysis reaction substrate bind to the active site of an enzyme.
in which ATP is converted to ADP
 Models of enzyme action Extremophile
 is a microorganism that thrives in extreme environments, environments in which humans and most other forms
1. Lock-and-Key Model (Emil Fischer) of life could not survive.
 only a substrate whose shape and chemical nature are  acidophiles - optimal growth at pH levels of 3.0 or below
complementary to those of the active site can interact  alkaliphiles - optimal growth at pH levels of 9.0 or above
with the enzyme.  halophiles - a salinity that exceeds 0.2 M NaCl needed for growth
 Hyperthermophiles- a temperature between 80°C and 122°C needed to thrive
 cryophiles - a temperature of 15°C or lower needed for growth
 piezophiles - a high hydrostatic pressure needed for growth

EXTREMOZYME
2. Induced-Fit Model (Daniel F. Koshland)  is a microbial enzyme active at conditions that would inactivate human enzymes as well as enzymes present in
 is a result of the enzyme’s flexibility other types of higher organisms.
 the enzyme active site, although not exactly  cellulases, amylases, xylanases, proteases, pectinases, keratinases, lipases, esterases, catalases, peroxidases and
complementary in shape to that of the substrate, is flexible enough that it phytases
can adapt to the shape of the substrate.  USE:
detergent formulations
the petroleum industry during oil well drilling operations
Enzyme specificity Extremophile: H. pylori and Stomach Ulcers
 the extent to which an enzyme’s activity is restricted to a specific substrate, a specific group of substrates, a • Helicobacter pylori, commonly called H. pylori, is a bacterium that
specific type of chemical bond, or a specific type of chemical reaction can function in the highly acidic environment of the stomach
 Absolute specificity • causes more than 90% of duodenal ulcers and up to 80% of gastric
• enzyme will catalyze only one reaction. ulcers.
• Ex. Catalase catalyzes the conversion of hydrogen peroxide (H2O2) to O2 and H2O. Hydrogen peroxide is • TREATMENT: acid-suppression or acid-neutralization medications
the only substrate it will accept PLUS
 Group specificity antibiotics(tetracycline, amoxicillin,metronidazole,clarithromycin,
• the enzyme will act only on molecules that have a specific functional group, such as hydroxyl, amino, or and levofloxacin).
phosphate groups. • Transmission: fecal–oral or oral–oral routes.
• Examples:
• Carboxypeptidase - it cleaves amino acids, one at a time, from the carboxyl end of a peptide
chain.
• Esterases- acts on ester bonds
• Peptidases-acts on peptide bonds
• Glycosidases- acts on glycosidic bonds.
 Linkage specificity
• the enzyme will act on a particular type of chemical bond, irrespective of the rest of the molecular
structure. This is the most general of the common specificities.
• Ex. Phosphatases hydrolyze phosphate-ester bonds in all types of phosphate esters a molecule closely resembling the a molecule that binds to a site on an a molecule that forms a covalent
 Stereochemical specificity substrate. Binds to the active site enzyme that is not the active site. The bond to a part of the active site,
• some enzymes are specific to only one isomer even if the compound is one type of molecule: and temporarily prevents normal substrate still occupies the active permanently preventing substrate
substrates form occupying it, thus site but the enzyme cannot catalyze the
• Ex. glucose oxidase catalyzes the oxidation of β-D-glucose but not α-D-glucose, and arginase catalyzes the
hydrolysis of L-arginine but not D-arginine. Maltase catalyzes the hydrolysis of α- but not β –glycosides. blocking the reaction. reaction due to the presence of the from occupying .
FACTORS THAT AFFECT THE RATE OF ENZYME ACTIVITY
inhibitor .
 Allosteric Enzymes TYPES OF CK
- Greek allo, means “other,” and stereos, means “site or space.”  muscle (CK-MM)
- is an enzyme with two or more protein chains (quaternary  brain (CK-BB),
structure) and two kinds of binding sites (substrate and regulator)  cardiac tissue (CK-MB) - important marker in the diagnosis of acute myocardial infarction (AMI)

Regulators- substances that bind at regulatory sites of allosteric TROPONIN

enzymes.  Description
• Troponin I and T are sensitive markers of cardiac injury.
Positive regulator increases enzyme activity; the shape of the active • Troponin I is found solely in the cardiac muscle, and
site is changed such that it can more readily accept substrate. • Troponin T is found in both cardiac and skeletal muscle.
 Clinical Significance
Negative regulator (a noncompetitive inhibitor; decreases enzyme • Troponin levels begin to rise within 4 hours of onset of chest pain. Levels should be drawn on admission
activity; changes to the active site are such that substrate is less and within 8 to 12 hours thereafter. Patients with elevated troponin levels are considered at high risk for a
readily accepted. significant cardiac event.
• Approximately 30% of patients with no elevation in CK-MB may demonstrate elevated troponin and thus be
diagnosed with a non-Q-wave myocardial infarction.

GASTROINTESTINAL TESTS
Alanine Aminotransferase/ serum glutamic pyruvic transaminase (SGPT).
REASON FOR REGULATION: Waste of energy  liver tissue. It is also located in myocardial, muscle, and renal tissue
 A cell that continually produces large amounts of an enzyme for which substrate concentration is always very  considered a specific marker for liver disease
low Aspartate Aminotransferase/ serum glutamic oxaloacetic transaminase (SGOT)
 A product of an enzyme-catalyzed reaction that is present in plentiful (more than needed) amounts.  found in the liver. It is also present in the heart, kidney, pancreas, lungs, and skeletal muscle
WAYS OF REGULATION  For diagnosis of liver disease
(1) feedback control associated with allosteric enzymes g-Glutamyl Transpeptidase
(2) production of enzymes in an inactive form: proteolytic enzymes and zymogens  an enzyme found in the liver, kidney, and pancreas. GGT levels are useful in the diagnosis and monitoring of
(3) covalent modification alcoholic liver disease
 Increased GGT may be seen in alcoholic liver disease, metastatic liver disease, obstructive jaundice,
Feedback Control associated with Allosteric enzyme cholelithiasis, and pancreatitis
a process in which activation or inhibition of the first reaction in a reaction sequence is controlled by a product of the Lactate Dehydrogenase
reaction sequence  enzyme involved in the interconversion of lactate and pyruvate.
 is found in many tissues, including heart, brain, liver, skeletal muscle, kidneys, lungs, and RBCs.
 LDH4 and LDH5 are present in liver tissue, and elevations may be seen in liver disease such as hepatitis and
cirrhosis.
 LDH1 and LDH2 may be useful in the diagnosis of myocardial infarction
Lipase
 enzyme that aids in the digestion of fat. It is primarily secreted by the pancreas.
proteolytic enzyme  useful in the diagnosis of pancreatitis and is considered a more specific marker for pancreatitis than amylase
 an enzyme that catalyzes the breaking of peptide bonds that maintain the primary structure of a protein. Amylase
zymogen or proenzyme  enzyme that aids in digestion by breaking down complex carbohydrates into simple sugars.
 is the inactive precursor of a proteolytic enzyme.  The majority of amylase is produced in the pancreas and salivary glands, and lesser amounts are secreted by the
EXAMPLES: fallopian tubes, lungs, thyroid, and tonsils
 Pepsinogen ------- pepsin  Serum amylase levels are most often used in the diagnosis of acute pancreatitis
 Tyrpsinogen -------trypsin
 Chymotrypsinogen ------- chymotrypsin
 Angiotensinogen ------- angiotensin CONDITION ISOENZYME PATTERN

Myocardial infarction Moderate elevation of LDH1, slight elevation of LDH2

BIOCHEMICAL IMPOTRTANCE: Enzymes
Important tool in diagnostic procedures (involving enzyme assay) Acute hepatitis Large elevation of LDH5, Moderate elevation of LDH4
 assist to know damaged tissues
 assist the extent of tissue damage Muscular dystrophy Elevation of LDH1-3
 helps to monitor the course of the disease
 used as a therapeutic means of diagnosing a vast array of diseases Megaloblastic anemia Large elevation of LDH1
Creatine kinase
 is an enzyme that is found primarily in skeletal and cardiac muscle and in smaller fractions in the brain Sickle cell anemia Moderate elevation of LDH1 and LDH2

Arthritis with joint infections Elevation of LDH5

 SERUM ENZYME MAJOR DIAGNOSTIC USE

Amylase Liver and pancreatic disease

Acid phosphate Prostate cancer

Alkaline phosphatase Liver and bone disease

Creatine phosphokinase Myocardial infarction and muscle disorders

Lactate dehydrogenase Myocardial infarction, leukemia, anemia

Renin Hypertension

Glutamic oxaloacetic transaminase Myocardial infarction

(SGOT)

Glutamic pyruvic transaminase Infectious hepatitis

(SGPT)

Trypsin Acute pancreatitis

Ceruplasmin Wilson’s disease