ENZYME

S
 ENZYMES
• An enzyme is a protein that acts as a catalyst for a
biological
reaction.
• Most enzymes are specific for substrates while enzymes
involved in digestion such as papain attack many
substrates
 ENZYMES
• Enzymes are complex organic catalysts of highly specific action that are of
vital importance in biological processes.
• They are manufactured by living cells, but are able to function
without the presence of these cells.
• They modify the speed of a reaction without being used up or appearing as
one of the end products of the reaction.
 NATURE OF ENZYMES:

• Enzymes are proteins of low molecular weight.

• They are coagulated and so become inactive by high temperature, alcohol, salts,
heavy
metals, and alkaloidal reagents.
• In general, they are soluble in water.
• Most reactions of living matter are accelerated by enzymes.
• Enzymes are responsible for the reactions like oxidation, reduction, hydrolysis, synthesis
and many others which are necessary during digestion, metabolism, respiration, energy
transfers and release.
ENZYMES ARE THE AGENTS
OF METABOLIC FUNCTION
regulatory enzymes capable of
sensing the momentary metabolic
needs of the cell and adjusting
their catalytic rates
accordingly.
 COFACTORS

• In addition to the protein part, many enzymes also have a nonprotein part
called a cofactor
• The protein part in such an enzyme is called an apoenzyme, and the
combination of apoenzyme plus cofactor is called a holoenzyme. Only
holoenzymes have biological activity; neither cofactor nor apoenzyme can
catalyze reactions by themselves
• A cofactor can be either an inorganic ion or an organic molecule,
called a coenzyme
• Many coenzymes are derived from vitamins, organic molecules that are
dietary requirements for metabolism and/or growth
 SOME C O M M O N ENZYMES A N D THEIR
ACTIVITIES
ENZYMES WHERE FOUND SUBSTRATE END PRODUCTS
I. Hydrolytic
Enzymes:
CARBOHYDRASES
Ptyalin or salivary amylase Saliva Starch & glycogen Maltose

Amylopsin or Pancreatic Pancreatic juice Starch & glycogen Maltose

Maltase Intestinal juice Maltose Glucose

Lactase Intestinal juice Lactose Glucose & galactose

Sucrase Intestinal juice Sucrose Glucose & fructose

ESTERASES OR LIPASES
Gastric Lipase Gastric juice in Emulsified fats Fatty acid & glycerol
Stomach
Streapsin or Pancreatic lipase Pancreatic juice Fats Fatty acid & glycerol

Phosphatases Tissues Hexophosphate Hexose & H3PO4

ENZYMES WHERE FOUND SUBSTRATE END PRODUCTS

PROTEASES:
. Pepsin Gastric juice Proteins Proteases &
Peptoses
Trypsin or Creatic Pancreatic juice Proteins, Polypeptides
Proteinase proteoses,
peptones
Pancreatic juice Proteins, Polypeptides
Chymotrysin proteoses,
peptones
Amino polypeptidases Intestinal juice Polypeptides Peptides &
& Carboxypeptidases amino acids

Dipeptidases Intestinal & Dipeptides Amino acids

Pancreatic juice
II. OXIDO-REDUCTASES:
Dehydrogenases Animal tissues Hydrogen donors Oxidized hydrogen
acceptor

Oxidases Plants & animal Carbohydrate proteins, fats in CO2 & H2O
Tissues tissues

Peroxidases Plant & animal Organic peroxides Oxygen

Tissues
. Catalases Plant & animal Hydrogen peroxides Water & oxygen
Tissues
III. FERMENTING ENZYMES:
. Zymase Yeast Monosaccharides CO2 and ethyl alcohol

Lactic acidase Lactic acid Lactose Lactic acid

Bacteria
IV. COAGULATING ENZYMES:
Rennin/Rennet Gastric juice Milk or casein Paracasein
 D EFINITION OF
TERMSor proenzyme-
• Zymogen is the inactive form of the enzymes. Ex. pepsinogen is
inactive form of pepsin; trypsinogen is the inactive form of the
trypsin.
• Activator or kinase- is a substance which activates the proenzyme. If the activator is an
organic substance it is called a kinase. Ex. HCl in the gastric juice activates pepsinogen
into pepsin, trypsinogen is activated by enterokinase in the intestinal juice into trypsin.
• Antienzymes- substances which inhibit enzyme activity. They are found in the lining of
the stomach and intestines, inhibit the action of pepsin on the stomach protein.
• Coenzymes- substance or electrolytes, or non-electrolytes which are required by
a
particular enzyme for its activity. Ex. Bile salts, chloride ions.
 HO W D O ENZYMES W ORK?
EXAMPLE: CITRATE
SYNTHASE
• Citrate synthase catalyzes a mixed Claisen condensation of acetyl CoA
oxaloacetate
and to give citrate
The Claisen condensation is a carbon–carbon bond forming reaction that
occurs between two esters or one ester and another carbonyl compound in
the presence of a strong base, resulting in a β-keto ester or a β-diketone.
• Normally Claisen condensation requires a strong base in an alcohol
solvent but citrate synthase operates in neutral solution
THE STRUCTURE OF CITRATE SYNTHASE
• D etermined by X -ray crystallography
• Enzyme is very large compared to substrates,
creating a complete environment for the reaction
 PROTEIN DENATURATION
• The tertiary structure of a globular protein is the result of
many intramolecular attractions that can be disrupted by
a change of the environment, causing the protein to
become denatured
• Solubility is drastically decreased as in heating egg
white, where the albumins unfold and coagulate
• Enzymes also lose all catalytic activity when
denatured
 U N D E R S T A N D I N G CATALYSIS

H O W D O ENZYMES
W ORK AS
C ATA LYSTS?
Lock-and-key model
portrays an enzyme as conformationally rigid
and able to bond only to substrates that exactly
fit the active site.
The INDUCED-FIT MODEL states a substrate binds to an active site and both
change shape slightly, creating an ideal fit for catalysis. Enzymes promote
chemical reactions by bringing substrates together in an optimal orientation, thus creating
an ideal chemical environment for the reaction to occur.
Substance which promotes
the activity of catalyst is
called “promotor” or
“activator”. it is also
called
“cataly for a
st such phenomenon
catalyst”.
catalyst is activatio
Examples n.
:
of Hydrogenation
oil to
vegetabl
vegetable ghee is
e
accelerated by
nickel.
FORMATION OF ENZYME-SUBSTRATE COMPLEX
 CLINICAL SIGNIFICANCE

• Today many competitive inhibitors are used as

drugs.
• Sulfanilamide, a sulfur drug, is an antibacterial
agent because it is a competitive inhibitor of an
enzyme- catalyzed reaction using p-aminobenzoic
acid in the synthesis of folic acid.
• Folic acid is a water-soluble vitamin.
Sulfanilamide is apparently not harmful to
man because folic acid is not synthesized
by man but obtained in the diet.
• Apparently bacteria can’t absorb sufficient
folic acid from the host and are
susceptible to inhibition by such drugs.
• Methotrexate (amethopterin) is a competitive
inhibitor of dihydrofolate reductase because
its structure is similar to dihydrofolate
• Methotrexate is used in cancer chemotherapy
because it blocks tetrahydrofolate synthesis
which is essential to the biosynthesis of the
thymine mononucleotide. Rapidly dividing
cancer cells which are synthesizing D N A are
more susceptible to methotrexate than slower
growing normal cells.
 ALTERNATIVE MEDICINE USING
Ratin ENZYMES Health
g Low back pain (chymotrypsin, Concerns
trypsin)
Pancreatic insufficiency (including
pancreatitis) Sprains and strains
(chymotrypsin, trypsin) Indigestion
(Lipase )
O steoarthritis (bromelain, trypsin,
rutosid combination)
Tendinitis (proteolytic enzymes)
Acne Rosacea
Chronic
candidiasis Food
allergies
Gastroesophageal
IN INDUSTRIES

• Cellulase is the major industrial

enzyme used to soften and fade
jeans (environment-friendly and
effective)
• http://www.europabio.org/documents/JEA
NS.p
df
 DAIRY PRODUCTS
INDUSTRIES
• Source: www.biotech.about.com.
• Biotechnology gives many industrial
applications that result in biotech products that
we use everyday at home
• Some of these are food science applications that
utilize enzymes to produce or make improvements
in the quality of different foods.
• In the dairy industry, some enzymes are required
for the production of cheeses, yogurt and other
dairy products, while others are used in a more
specialized fashion to improve texture or flavour
 CHEESE-MAKING

• Cheese consists of proteins and

fat from milk, usually the milk of
cows, buffalo, goats, or sheeps. It
is produced by coagulation of
the milk protein casein.Typically,
the milk is acidified and addition
of the enzyme rennet causes
coagulation.
•
COENZYMES
 COEN ZYMES
• There are other groups that contribute to the reactivity of
enzymes beside amino acid residues.
• These groups are called cofactors - chemicals required
by apoenzymes (inactive) to become holoenzymes
(active).
• There are two types of cofactors:
1) essential ions - metal ions -inorganic
2)coenzymes - organic molecules that act as group-transfer
reagents (accept or donate groups)- can also be H + and/or e-
Both provide reactive groups not found on a.a. side chains.
1) METABOLITE COENZYMES

• synthesized by common metabolites

• include nucleoside triphosphates
• most abundant is ATP, but also include uridine diphosphate glucose (UDP-
glucose) and S-adenosylmethionine
• ATP can donate all of its three phosphoryl groups in group-transfer
reactions
• S-adenosylmethionine (SAM) can donate its methyl group in biosynthetic
reactions.
• UDP-glucose is a source of glucose for synthesis of glycogen in animals and
starch in plants.
2) vitamin-derived coenzymes
• Vitamins are required for coenzyme synthesis and must be supplied in
the diet
• Lack of particular vitamins causes disease
• There are two catagories of vitamins:
1)water-soluble - B vitamins and vit. C- required daily in diet,
excess excreted in urine
2) lipid-soluble - vitamins A, D,
E, K Intake must be limited
Stored in fat
 B VITAMINS A N D THEIR C O E N Z Y M E S VITAMINB3
NIACIN

nicotinic acid  nicotinamide

• Get niacin in enriched cereals, meat, legumes.
• N A D + and NADP + are the coenzymes (cosubstrates).
• N A D + consists of 2 5’ribonucleotides (AMP and nicotinamide monomucleotide) joined by
a phosphoanhydride linkage.
• For NADP + , have a phosphoryl group on 2’-oxygen.
• Both coenzymes act as cosubstrates for dehydrogenases  catalyze the oxidation of
substrates by transfer
of 2e- and 1H + N A D H and N A DPH.
 VITAMIN B 2 (RIBOFLAVIN)

• Coenzymes are flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD).
• Riboflavin found in milk, whole grains, liver.

• The coenzymes serve as prosthetic groups involved in 1e- or 2e- transfers.

FAD + 2e- + 2H +  FADH 2

FMN + 2e- + 2H +  FMNH 2

• Enzymes that require FAD or FMN are called flavoenzymes or flavoproteins.

• Can actually donate 1 or 2 e- at a time  form partially oxidized compound when only 1e- is donated 
relatively stable.
 VI TAMI N B 1 (THIAMINE)

• Structure: pyrimidine ring and positively charged thiazolium ring.

• Found in husks of rice and other cereals, liver, meat, particularly
pork.
• Deficiency in thiamine causes beriberi - extensive nervous
system
and circulatory system damage, muscle wasting, edema.
• Coenzyme form is thiamine pyrophosphate (TPP) -
synthesized by transfer of pyrophosphoryl group from ATP via
thiamine pyrophosphate synthetase.
• Used primarily in decarboxylases as a coenzyme.
 V ITAMI N B6 FAMILY
• pyridoxine, pyridoxal, pyridoxamine are the vitamins.
• Act as prosthetic groups.
• Formed by the following reaction:
pyridoxine + ATP  pyridoxine 5’phosphate  pyrodoxal 5’ phosphate
(PLP).
• Lack of B6 results in defects in protein metabolism.
 B IO T I N

• Synthesized by intestinal bacteria.

• Prosthetic coenzyme is called biocytin - covalently linked to Lys residue
in active site.
• Involved in carboxyl group transfer reactions and ATP-
dependent carboxylations.
• E.g. pyruvate carboxylase
pyruvate + HCO 3 oxaloacetate
- 

• Binds to HC O 3- and acts as a C O 2 carrier

FOLI C A C I D O R
FOLATE
• Found in green leafy vegetables, liver, yeast.
• Coenzyme form is tetrahydrofolate.
• Used by enzymes that transfer 1-C units as methyl groups (CH 3-).
• Another folate coenzyme is tetrahydrobiopterin - used in
hydroxylases
 VITAMI N B5: P A N T H O T H E N I C
ACID

• Used in coenzyme A formation.

• Reactive center is -SH group
• Key in all acyl-group transfers
• Coenzyme form is phosphopantethine - added to
serine residue of protein --> acyl carrier protein
(ACP) --> important in fatty acid synthesis.
 VITAMIN B12 O R C O B A L A M I N

• Found in organ meat (kidney and liver).

• It is a prosthetic coenzyme.
• Ring structure similar to heme, with cobalt atom in center.
• Involved in molecular rearrangements.
• Deficiency in B12 results in pernicious anemia (decreased production of blood cells
from bone marrow).
VITA M I N C O R A S C O R B I C
ACID

• Found in fresh fruit and vegetables.

• Participates in hydroxylation reactions, e.g. collagen
synthesis.
• Deficiency causes scurvy.
 LIPID V ITA M I N S /VITA M I N A O R
RETIN O L
• Is a 20 carbon lipid molecule.

• Found in carrots, yellow vegetables, liver, egg yolk, milk

products. carotene  vitamin A
Exists in three forms:
1) retinol and 2) retinoic acid - bind to intracellular protein
receptors  regulates
gene expression
2) retinal - prosthetic group of rhodopsin
 VITAMI N D

• Exists as several lipids;

• 1) D3 - made in skin exposed to sunlight.
• 2) D2 - additive in fortified milk
• Deficiency causes rickets in children or osteomalacia
in adults --> insufficient Ca phosphate deposition in
bone.
VITA M I N E O R A -
TO C O P H E RO L

• an antioxidant that scavenges free

radicals.
 VITAMI N K O R P H Y L L O Q U I N O N E

• Found in plants.
• Required for synthesis
of proteins involved in
blood coagulation.
 UBIQUINONE OR COENZYME Q

• Ring with hydrophobic tail --> inserted into

membranes.
• Transports e- between enzyme complexes in
inner mitochondrial membrane.
• Related molecule is plastiquinone - found in
thylacoid membrane of chloroplasts.
CYTOCHROMES

• Hemo-containing protein coenzyme Fe3+ <-->

Fe2+.
• Classified as a, b, c based on absorption spectra.
• Transfers e-.
Explain the mechanism of
Ubiquinone or coenzyme
Q10 supplement