Chapter 4: Enzymes and Vitamins - Typical metal ion cofactors - Zn2+, Mg2+,

Mn2+, and Fe2+

Enzymes are catalysts and are not consumed in
- Nonmetallic ion cofactor - Cl-
the reactions
- proteins that act as a catalyst for biochemical - Inorganic ion cofactors derived from dietary
reactions minerals
- human body has 1000s of enzymes
- most effective catalysts known Nomenclature and Classification of
- most enzymes are globular proteins Enzymes:
- A few enzymes are now known to be ribonucleic
acids (RNA) Nomenclature: Most commonly named with
- undergo all the reactions of proteins including reference to their function
denaturation
- Type of reaction catalyzed
Enzyme activity is dramatically affected by: - Identity of the substrate
- Alterations in pH
- Temperature A substrate is the reactant in an enzyme
- Other protein denaturants catalyzed reaction:
- is the substance upon which the enzyme
Enzyme Structure: Simple and “acts.”
Conjugated Enzymes
- E. g., In the fermentation process sugar to
2 TYPES OF ENZYMES be converted to CO2, therefore in this reaction
sugar is the substrate
1. Simple enzymes composed only of protein Three Important Aspects of the
(amino acid chains)
Naming Process
2. Conjugated enzymes has a nonprotein part 1. Suffix -ase identifies it as an enzyme
in addition to a protein part.
- E.g., urease, sucrase, and lipase are all
Apoenzyme: Protein part of a conjugated enzyme designations
enzyme.
- Exception: The suffix -in is still found in the
A cofactor: Nonprotein part of a conjugated
names of some digestive enzymes, E.g.,
enzyme.
trypsin, chymotrypsin, and pepsin
A holoenzyme is the biochemically active
conjugated enzyme 2. Type of reaction catalyzed by an enzyme is
Apoenzyme + cofactor = holoenzyme often used as a prefix
(conjugated enzyme)
- Oxidase - catalyzes an oxidation reaction
Cofactors - Hydrolase - catalyzes a hydrolysis reaction
- are important for the chemically reactive
enzyme 3. Identity of substrate is often used in
- are small organic molecules or Inorganic addition to the type of reaction
ions
- E.g. Glucose oxidase, pyruvate carboxylase,
- Organic molecule cofactors: also called as
and succinate dehydrogenase
co-enzymes or co-substrates
- Co-enzymes/co-substrates are derived Function of the following enzymes.
from dietary vitamins
Maltase- hydrolysis of maltose
- Inorganic ion cofactors
Lactate dehydrogenase- removal of hydrogen - The reaction involves addition of a water
from lactate ion molecule to a bond to cause bond breakage
- Hydrolysis reactions are central to the
Fructose oxidase- oxidation of fructose process of digestion:
Maleate isomerase- rearrangement - Carbohydrases hydrolyze glycosidic
(isomerization) of maleate ion bonds in oligo- and polysaccharides (see
reaction below)
Six Major Classes - Proteases effect the breaking of peptide
linkages in proteins
Enzymes are grouped into six major classes - Lipases effect the breaking of ester
based on the types of reactions they catalyze linkages in triacylglycerols

Class Reaction Catalyzed • Lyase- is an enzyme that catalyzes the

Oxidoreductases Oxidation- reductions addition of a group to a double bond or the
Transferases Functional group removal of a group to form a double bond in
transfer reaction a manner that does not involve hydrolysis or
Hydrolases Hydrolysis reaction oxidation
Lyases Reactions involving
addition or removal of
groups form double
• - Dehydratase: effects the removal of the
components of water from a double bond
bonds
Isomerase Isomerization
reactions
• - Hydratase: effects the addition of the
components of water to a double bond
Ligases Reactions involving
bond formation
coupled with ATP
• Isomerase and Ligase
hydrolysis
• Isomerase is an enzyme that catalyzes the
isomerization (rearrangement of atoms)
Oxidoreductase Enzyme- catalyzes an
reactions.
oxidation–reduction reaction:
- Oxidation and reduction reactions are
always linked to one another
• Ligase is an enzyme that catalyzes the
formation of a bond between two molecules
- An oxidoreductase requires a coenzyme
involving ATP hydrolysis:
that is either oxidized or reduced as the
- ATP hydrolysis is required because such
substrate in the reaction.
reactions are energetically unfavorable
- E.g., Lactate dehydrogenase is an
- Require the simultaneous input of energy
oxidoreductase and the reaction catalyzed
obtained by a hydrolysis of ATP to ADP
is shown below

Transferase- is an enzyme that catalyzes MODELS OF ENZYME ACTION

the transfer of a functional group from one
molecule to another • Enzyme Active Site

Two major subtypes:

• The active site: Relatively small part of an
enzyme’s structure that is actually involved
Transaminases - catalyze transfer of an in catalysis:
amino group to a substrate
Kinases - catalyze transfer of a phosphate • - Place where substrate binds to enzyme
group from adenosine triphosphate (ATP) to - Formed due to folding and bending of the
a substrate protein.
Hydrolase- is an enzyme that catalyzes a - Usually a “crevice like” location in the
hydrolysis reaction enzyme
- Some enzymes have more than one active Group Specificity:
site - Involves structurally similar compounds
that have the same functional groups.
Enzyme Substrate Complex - E.g., Carboxypeptidase: Cleaves amino
- Needed for the activity of enzyme acids one at a time from the carboxyl end of
- Intermediate reaction species formed the peptide chain

•
when substrate binds with the active site
- Orientation and proximity are favorable Linkage Specificity:
and reaction is fast - Involves a particular type of bond
irrespective of the structural features in the
Two Models for Substrate Binding to vicinity of the bond
Enzyme - most general of enzyme specificities
- E.g., Phosphatases: Hydrolyze
Lock-and-Key model: phosphate–ester bonds in all types of
- Enzyme has a pre-determined shape for phosphate esters
the active site
- Only substrate of specific shape can bind FACTORS THAT AFFECT ENZYME
with active site ACTIVITY
Induced Fit Model:
- Substrate contact with enzyme will change
• Temperature
- Higher temperature results in higher
the shape of the active site kinetic energy which causes an increase in
- Allows small change in space to number of reactant collisions, therefore
accommodate substrate (e.g., how a hand there is higher activity.
fits into a glove) - Optimum temperature: Temperature at
which the rate of enzyme catalyzed reaction
Forces That Determine Substrate is maximum
Binding - Optimum temperature for human enzymes
-H-bonding is 37ºC (body temperature)
- Hydrophobic interactions - Increased temperature (high fever) leads
-Electrostatic interactions to decreased enzyme activity

ENZYME SPECIFICITY • pH
- pH changes affect enzyme activity
Absolute Specificity: - Drastic changes in pH can result in
- An enzyme will catalyze a particular denaturation of proteins
reaction for only one substrate - Optimum pH: pH at which enzyme has
- This is most restrictive of all specificities maximum activity
(not common) - Most enzymes have optimal activity in the
- E.g., Urease is an enzyme with absolute pH range of 7.0 - 7.5
specificity - Exception: Digestive enzymes
- Pepsin: Optimum pH = 2.0
Stereochemical Specificity: - Trypsin: Optimum pH = 8.0

•
- An enzyme can distinguish between
stereoisomers. Substrate Concentration
- Chirality is inherent in an active site (amino - At a constant enzyme concentration, the
acids are chiral compounds) enzyme activity increases with increased
- L-Amino-acid oxidase - catalyzes reactions substrate concentration.
of L-amino acids but not of D-amino acids. - Substrate saturation: the concentration at
which it reaches its maximum rate and all of
the active sites are full
- Turnover Number: Number of substrate Examples: Heavy metal ions Pb2+, Ag+, and
molecules converted to product per second Hg2+.

•
per enzyme molecule under conditions of
optimum temperature and pH Irreversible Inhibition
- An irreversible enzyme inhibitor inactivates
Enzyme Concentration: enzymes by forming a strong covalent bond
- not consumed in the reactions they with the enzyme’s active site.
catalyze - The structure is not similar to enzyme’s
- At a constant substrate concentration, normal substrate
enzyme activity increases with increase in - The inhibitor bonds strongly and
enzyme concentration increasing substrate concentration does not
- The greater the enzyme concentration, the reverse the inhibition process
greater the reaction rate. - Enzyme is permanently inactivated.
- E.g., Chemical warfare agents (nerve
ENZYME INHIBITION gases) and organophosphate insecticides

Enzyme Inhibitor: a substance that slows REGULATION OF ENZYME

down or stops the normal catalytic function ACTIVITY
of an enzyme by binding to it. - Cellular processes continually produces
large amounts of an enzyme and plentiful
Competitive Inhibitors: Compete with the amounts of products if the processes are
substrate for the same active site not regulated.
- Will have similar charge & shape - General mechanisms involved in
regulation:
Noncompetitive Inhibitors: Do not - Proteolytic enzymes and zymogens
compete with the substrate for the same covalent modification of enzymes
active site - Feedback control Regulation of enzyme
- Binds to the enzyme at a location other activity by various substances produced
than active site within a cell
- The enzymes regulated are allosteric
Reversible Competitive Inhibition enzymes
- A competitive enzyme inhibitor: resembles
an enzyme substrate in shape and charge
- Binds reversibly to an enzyme active site
• Properties of Allosteric Enzymes
All allosteric enzymes have quarternary
and the inhibitor remains unchanged (no structure:
reaction occurs) - Composed of two or more protein chains
- The enzyme - inhibitor complex formation
is via weak interactions (hydrogen bonds, • Have at least two of binding sites:
etc.). - Substrate and regulator binding site
- Competitive inhibition can be reduced by
simply increasing the concentration of the Active and regulatory binding sites are
substrate. distinct from each other:
- Located independent of each other
Reversible Noncompetitive Inhibition - Shapes of the sites (electronic geometry)
- A noncompetitive enzyme inhibitor are different
decreases enzyme activity by binding to a
site on an enzyme other than the active site.
• Binding of molecules at the regulatory site
causes changes in the overall three
- Causes a change in the structure of the
dimensional structure of the enzyme:
enzyme and prevents enzyme activity.
- Increasing the concentration of substrate
does not completely overcome inhibition.
- Change in three dimensional structure of the - Removal of the phosphate group
enzyme leads to change in enzyme activity (dephosphorylation) catalyzed by a
- Some regulators increase enzyme activity – phosphatase enzyme.
activators - Phosphate group is added to (or removed
- Some regulators decrease enzyme activity – from) the R group of a serine, tyrosine, or
inhibitors threonine amino acid residue in the enzyme
regulated.
Feedback Control
- A process in which activation or inhibition ANTIBIOTICS THAT INHIBIT ENZYME
of the first reaction in a reaction sequence is ACTIVITY
controlled by a product of the reaction
sequence. - An anitibiotic is a substance that kills
- Regulators of a particular allosteric bacteria or inhibits their growth
enzyme may be: - Antibiotics usually inhibit specific enzymes
- Products of entirely different pathways of essential to life processes of bacteria
reaction within the cell - Two families of antibiotics considered in
- compounds produced outside the cell this discussion are sulfa drugs and
(hormones) penicillins

Proteolytic Enzymes and Zymogens • Sulfa Drugs

- 2nd mechanism of regulating enzyme - Many derivatives of sulfanilamide
activity: collectively called sulfa drugs exhibit
- Production of enzymes in an inactive antibiotic activities
forms (zymogens) - Sulfanilamide is structurally similar to
- Zymogens are “turned on” at the PABA (p-aminobenzoic acid)
appropriate time and place - Many bacteria need PABA to produce
Example: proteolytic enzymes: Most coenzyme, folic acid
digestive and blood-clotting enzymes are - Sulfanilamide is a competitive inhibitor of
proteolytic enzymes enzymes responsible for converting PABA
- Hydrolyze peptide bonds in proteins to folic acid in bacteria
- Proteolytic enzymes are generated in an - Folic acid deficiency retards bacterial
inactive form and then converted to their growth and that eventually kills them
active form - Sulfa drugs don’t affect humans because
we absorb folic acid from our diet
Covalent Modification of Enzymes
- 3rd Mechanism for regulation of enzyme • Penicillins
activity - Accidently discovered by Alexander
- Covalent modification: A process in which Fleming in 1928
enzyme activity is altered by covalently - Several naturally occurring penicillins and
modifying the structure of the enzyme: numerous synthetic derivatives have been
- Involves adding or removing a produced
group from an enzyme - All have structures containing a four-
membered Beta-lactam ring fused with a
-Most common covalent modification: five-membered thiazolidine ring
addition and removal of phosphate group: - Selectively inhibits transpeptidase by
- Phosphate group is often derived covalent modification of serine residue
from an ATP molecule. - Transpeptidase catalyzes the formation of
- Addition of the phosphate peptide cross links between
(phosphorylation) catalyzed by a polysaccharides strands in bacterial cell
Kinase enzyme walls
Cipro - Co-substrate in the formation of structural
- The antibiotic ciprofloxacin hydrochloride protein collagen
(Cipro for short) - Involved in metabolism of certain amino
- Considered the best broad-spectrum acids
antibiotics because it is effective against -100 mg/day saturates all body tissues -
skin and bone infections as well as against Excess vitamin is excreted
infections involving the urinary,
gastrointestinal, and respiratory systems RDA (mg/day):
- It is the drug of choice for treatment of - Great Britain: 30
traveler’s diarrhea - United States and Canada: 60
- Bacteria are slow to acquire resistance to - Germany: 75

•
Cipro.
- Biochemical threats associated Vitamin B
with terrorism has thrust Cipro into the The preferred and alternative names for the
spotlight because it is effective against B vitamins:
anthrax. - Thiamin (vitamin B1)
- Riboflavin (vitamin B2)
MEDICAL USES OF ENZYMES - Niacin (nicotinic acid, nicotinamide, vitamin
B3)
Diagnose certain diseases: - Vitamin B6 (pyridoxine, pyridoxal,
- Enzymes produced in certain organ/ pyridoxamine)
tissues if found in blood may indicate certain - Folate (folic acid)
damage to that organ/tissue - Vitamin B12 (cobalamin)
- Pantothenic acid (vitamin B5)
GENERAL CHARACTERISTICS OF - Biotin
- Exhibit structural diversity
VITAMINS - Major function: B Vitamins are
- Organic compounds components of coenzymes
- Must be obtained from dietary sources
- Human body can’t synthesize in enough
amounts FAT SOLUBLE VITAMINS
- Essential for proper functioning of the body
- Needed in micro and milligram quantities • Vitamins A, D, E, K
- 1 Gram of vitamin B is sufficient for - Involved in plasma membrane processes
- More hydrocarbon like with fewer
500,000 people
- Enough vitamin can be obtained from functional groups
balanced diet
- Supplemental vitamins may be needed
• Vitamin A
- Has role in vision - only 1/1000 of vitamin
after illness A is in retina
- Many enzymes contain vitamins as part of - 3 Forms of vitamin A are active in the body
their structures - conjugated enzymes - Derived from b-carotine
- Two Classes
- Water Soluble and Fat Soluble Functions of Vitamin A
- Synthetic and natural vitamins are same Vision: In the eye- vitamin A combines with
- 13 Known vitamins opsin protein to form the visual pigment
rhodopsin which further converts light
energy into nerve impulses that are sent to
WATER SOLUBLE VITAMINS the brain.
Vitamin C Regulating Cell Differentiation - process in
- Humans, monkeys, apes and guinea pigs which immature cells change to specialized
need dietary vitamins cells with function.
Examples: Differentiation of bone marrow S
cells white blood cells and red blood cells.

- Maintenance of the healthy of epithelial

tissues via epithelial tissue differentiation.
- Lack of vitamin A causes such surfaces to
become drier and harder than normal.

Reproduction and Growth: In men, vitamin

A participates in sperm development. In
women, normal fetal development during
pregnancy requires vitamin A.

Vitamin D
- Two forms active in the body: Vitamin D2
and D3
- Sunshine Vitamin: Synthesized by UV light
from sun
- It controls correct ratio of Ca and P
bone mineralization (hardening)
- As a hormone it promotes Ca and P
absorption in intestine

Vitamin E
- Four forms of Vitamin Es: a-, b-, g- and d-
Vitamin E
- Alpha-tocopherol is the most active
biological active form of Vitamin E
- Peanut oils, green and leafy vegetables
and whole grain products are the sources of
vitamin E
- Primary function: Antioxidant – protects
against oxidation of other compounds

Vitamin K
- Two major forms; K1 and K2
- K1 found in dark green, leafy vegetables
- K2 is synthesized by bacteria that grow in
colon

Dietary need supply: ~1/2 synthesized by

bacteria and 1/2 obtained from diet

Active in the formation of proteins involved

in regulating blood clotting
 VITAMINS: LECTURE - daily requirement in adults is 5000
units USP.
VITAMINS -
- are organic compounds, occurring naturally in VITAMIN D
foods, which are required in minute - precursor of a hormone, which regulates
amounts for normal growth, maintenance calcium and phosphate metabolism.
and reproduction.
- there are 2 important types of vitamin D,
- differ with some other organic foodstuffs
because they do not enter into the tissue 1.Dehydrocholesterol from the skin, if
structure and do not undergo degradation irradiated becomes cholecalciferol which is
for purposes of providing energy. also known as Vit. D3.
2.Ergosterol from plants, if irradiated
CLASSIFICATION becomes ergocalciferol which is also known
1.FAT SOLUBLE VITAMINS as Vit. D2.
Vitamin A
Vitamin D
Vitamin E - deficiencies here in the Phil. is very rare
Vitamin K because of its abundance and if there is any
2.WATER SOLUBLE VITAMINS these are rachitic children, bowlegs and
knock-knees.
Vitamin C
- also known as anti-rachitic vitamin
Vitamin P
- dosage for pregnant and nursing
Vitamin B-complex
women is 500-800 units.

VITAMIN E
FAT SOLUBLE VITAMINS
VITAMIN A - unsaturated lipids coming from both plants
- there are 2 sources, retinal (aldehyde) from and animals such as germ oils and carotene
animals and retinol (alcohol) from plants. in yellow plants.
- retinal is an active component of rhodopsin - protects the lipids of the cell membrane
(visual purple). from the destructive effect of oxygen.
- provitamin A is from beta-carotene of plants - anti-oxidants for both plants and animals.
and is converted to Vitamin A by
- also known as alpha tocopherol or Vit. X.
oxidation.
- deficiencies:
VITAMIN K
- degenerative changes in the skin - component of a carboxylating enzyme.
and mucous membrane including those of
glandular structures, making them - synthesizes prothrombin to thrombin in the
susceptible to infection, thus known an anti- liver, essential to blood clotting.
infective vitamin.
- deficiency of rhodopsin is - deficiency results in the failure of blood to
NYCTALOPIA OR NIGHT BLINDNESS clot.
(ability of the eyes to adapt itself to vision in
- also known as anti-hemorrhagic vitamin.
dim light after exposure to bright light).
WATER SOLUBLE VITAMINS: - required for methylation in the body.
5. Niacin or Nicotinic Acid or
Nicotinamide or Vit. B3
VITAMIN P (CITRIN & RUTIN)
- pellagra preventive factor or P-P
- influence capillary fragility and permeability.
factor.
-
- citrin is isolated from lemon peel. 6. Pyrodoxine or Vit. B6
- anti-dermatitis factor.
- rutin has been found in cured tobacco,
leaves and tops of green buck-wheat. - anti-acrodynia factor.
7. Biotin or Vit. B7. or Vit. H
VITAMIN C - also known as coenzyme R.
- sources are citrus fruits, tomatoes, cabbage
and spinach. - anti-eggwhite injury factor.

- formation of intercellular material. - necessary for mold, bacteria and yeast

growth.
- deficiency causes scurvy in man.
8. Inositol or Vit. B8
- also known as ASCORBIC ACID. - mouse anti-allopecia factor.
- cure spectacle eyes in rats.

VITAMIN – B COMPLEXES 9. Folic Acid or Vit. B9 or Vit Bc.

- anti-anemic factor in man.

1. Thiamine or Vit. B1. - also known as vit. M.

- heat –labile factor.
- also known as PGA
- anti-neuritic factor. (Pteroylglutamic acid).

- anti-beri-beri. 10. Cyanocobalamin or Vit. B12.

- anti-anemic vitamin.
- Aneurin.
- anti-pernicious anemia factor.
2. Riboflavin or Vit. B2 or Vit. G.
11. PABA (Para Amino benzoic Acid)
- growth factor. or Vit. Bx.
- form part of the folic acid molecule.
- lactoflavin.
- anti gray hair factor.
3. Pantothenic Acid or Vit. B5.

- filtrate factor.
- chick dermatitis factor.
4. Choline or Vit. B4
- growth factor.
- prevents perosis (shortening and
thickening of bones in chicks).