Enzymes

Biological catalysts .

RNA
are composed of , eg peptidyl transferase
{ are globular There is a grp of enzymes called ribosomes which
. .

.
Most cntymes are made of protein .

enable metabolic rxns within living organisms to proceed

metabolic rxns critical to sustaining life They are innit because they rapidly at low temp
Enzymes catalysed wide range
of
.
•

'

Ehlymes also ensure that metabolism (both anabolic & catabolic vxns ) proceeds stepwise in an orderly fashion .

Anabolic rxns : Synthesis Of Molecules from smaller molecules and usually require ATP . Usually involves condensation ( Eg synthesis of polysaccharide )
-

>
Catabolic rxns : Breakdown of molecules & Usually release energy stored in the bonds . Often involved oxidation or hydrolysis (
Eg Hydrolysis of glycogen
.

Properties of enzymes
1. Enzymes are highly specific in their action
-
Most enzymes act on only one type of substrate molecule , while others act
on group of highly similar
substrate molecules Mode Of Action Of Enzymes Rate Of enzyme
.

2. Enzymes are effective in small amounts

rxn
catalysed
A very
.

small amount of catalyst can bring about the conversion of a large 1 Active site
-

amount of substrate This is because enzymes remain chemically unchanged

.

at the end of the rxns they catalyse , { therefore can be reused &
.
.
The active site is a depression on the surface of the enzyme where substrates enter bind to Inhibitors
3. Enzymes are very efficient enabling .
rxns to proceed 10 ' -108 times faster { Where biological rxns occur .

than Un catalysed rxns They are said to have high turnover no The no Ot
a . -

} distribution of electrical charge which only

.

substrate molecules converted into products by one molecule of enzyme in

-
The active site has a specific 3D conformation u ✓
one second When the enzyme is
-

fully saturated with substrate . Enlymes speed substrates of complementary shape & charge will fit into lspdtidl fit { chemical fit
respectively) , Temperature
✓
✓ substrate conc
up the rates of both the forward { backward rxns . hence giving rise to the
specificity of enlyme action PH Enzyme conc
Rate of enzyme activity
.

^
>
4. The presence of enzymes does not alter the nature or properties of the end pelts ghost / Max Most eniymesfn within A. A -110W enlyme conc , the rate of A. At 10W substrate cone , the rate of rxn
a narrow pit range
-
-

The amino acid residues in have different roles optimum

.

an
'

enzyme
.

of the rxn temp rate otrxn rxn is low As eniymeconc increases increases with substrate conc as many Non-competitive inhibitors
Rdteofalllme activity
.
.
,
or
.

of the rate of rxn Increases linearly This

Responsible for Cdtaly sing the chemical
rate
ptimumpt, .

enzyme molecules have unoccupied active

Catalytic residues bonds in the substrate which enzyme
& rxn eg
: act On
5. Enzymes are soluble Is because sites Substrate conc is the limiting factor
U
.

in water work in the aqueous environment within activity as enzyme conc increases
.

living will be broken Pin rate

.

more active sites are now available for

. .

As substrate conc T the Vdteotrxn T

.

Cells otrxn
-

Competitive Inhibitors
,
successful collisions with the substrate
.

This is because there are more

.

residues : Ensure the specific conformation of the active site

frequency of
.

contact .

There is an increase in the linearly . ,

6 . The activity of enzymes is attested by temperature pH , enzyme conc & : Hold substrate in the correct orientation & position successful collisions b/w the substrate substrates available for successful collisions
A COMBO Und has no structural similarity to the
} with the enzymes There is an 1h in the
, -

the presence of activators inhibitors and enzyme , increasing the rate of formation
substrate { binds to the enzyme at a site other than the
.
.

Structural residues : C the rest of the residues within the enzyme ] work to maintain the overall > Temperature 1°C of E S complexes , thus
frequency of successful collisions btw
, increasing
the rate of A compound that is structurally similar to
-

C. ← ☐
.

AL B → →
formation of products Enzyme
-

globular shade of the molecule .

conc is . the the substrate } entry me M rate of formation the actual active site Canoe steric site ) , resulting in a decreased rxn
substrate and can enter & bind
.

to
limiting factor of E S complexes thus µ the rate of formation
,
> PH -

← ☐ → A← ☐ → rate is known as
.

a non competitive inhibitor

site the enzyme is known
,
the
-

A. At low temperatures near or below 0°C active of as competitive .

Effect limiting fdc

,

The catalytic and contact residues make up the active site These amino acids are often some of pH the rate Ot enzyme of products Substrate is the
on
activity B. At very high enzyme conc , enzyme cone
•

as
enzyme activity is very low The enzyme Is said
. .

Inhibitor
. .

class Type of rxn catalysed inactivated

.

distance apart in the polypeptide chain but are brought into close proximity by the folding
.

of increases , the rate of rxn remains Upon binding

•

to be as it possesses minimal conc of the noncompetitive Inhibitor

-

the chain kinetic energy for collision

. w substrate to
A. At optimum pH maximum rate otrxn occurs , An A to the allosteric site the enzymes 's 3D conformation
Catalyses the transfer of phosphate grps from ATP to specific constant In enzyme conc will not
-

When the competitive inhibitor is bound at the ,

form
> . .

E S complexes as bonds the secondary { tertiary B- At very high substrate cone > as substrate 00h0
maintaining enzyme active site it prevents the substrate from is such that the conformation of its active site
-

substrates in a process known as

phosphorylation further * in the changed
.

. .

The substrate enters } binds to the active site to form an enzyme substrate
-

complex structures of the eniyme are intact , enabling

result in a rate of rxn .

T , the rate Ot remains constant This is because

,

formation of
kinds es The addition of phosphates often changes a protein from the
rxu the site the
limiting entering preventing altered { longer bind to the enzyme
.

( non eniyme { substrate molecules )

B. As temperature increases before optimum Enzyme conc is no longer the , is the substrate can no
covalent complex of
highest frequency
,
inactive form C conformational change ) These active proteins can of successful
-

collisions sites are saturated with & formation

temperatures is reached the rate of mint all the avail enlyme active E- s complexes of products hence decreasing
.

btw substrate and enzyme This increases the factor

,
in turn activate
,
proteins , triggering
,
other numerous runs in a cell ✓
The increase in heat lathe kinetic energy rate of formation of E- s complexes , thus
-
, substrate conc is the .

limiting substrate, at any one time .

( any extra substrate has to
the rate of
active site .

at once .

Temporary weak bonds that hold

of the substrate and enzyme The frequency factor until the product released from the enzyme active
rxn .

catalyse the removal of phosphate groups from their substrates { substrate tgt the of formation of products wait
.

enzyme increasing
.

rate
Rate of yxn { Formation Of Enlyme
. .

the eteect of s of successful collisions btw the substrate before active site for r'M inhibitor complexes prevents the
kinases
Phosphatases ← bind
-

c reverse site it can enter the

-

to
Enwgme inhibitor complexes are formed , because
-

,
a
of E S complexes { prevents the
of a protein often deactivates the protein
-

The dephosphorylation .
to occur . ) substrate longer the
conc is no
limiting fac .

the competitive inhibitor competes with the formation -

formation
C. At optimum temperature , Max rate of rxns B. At DH other than optimum pH , the rdtlotr 'M -
is the limiting fac of products the rate otrxn
live the transfer 0TH - 0
decreasing
or
catalyse oxidation or reduction rxns
2. Activation energy Ea enzyme conc
.

substrate for binding at the enzyme active

.

At the optimum temperature , there the change in PH

.

occurs
decreases This is because
,
one molecule to another in redox vxns ] At nigh adding more
.

from
,
enzyme conc,
.

Oxidoreductases electrons
,

frequency
, .

Every chemical rxn involves bond

breaking and bond forming ishighest of successful eniymeshasnoettectonrateotrxn
eg dehydrogenases which involved in respiration alters the ionic charge of the acidic ( COO I
are
.
- .

collisions btw substrate h enzyme

.

very high
.

Date otvxn is limited by substrate Even at substrate conc , the Mdx rate of rxn
-

.
cone .

Transfer a functional grp to specific substrates , eg DNA .

-
The initial investment of
energy before a rxn can occurs is known as the activation energy ( Ea ) E. basic CNH } -1112 groups on the amino acids at -

At 10W Substrate cone as cone of substrate , } in the presence of the non competitive inhibitor
- is lower
Transferases the active site of the enzyme The ionic bonds -

inhibitor are similar the competitive ihibitor

methyltransferase Enzymes
,
than the maximum rate of rxh
to proceed faster by lowering As enzyme comet , adding in the absence of the
.

enable rxns the activation energy and hydrogen bonds that help to maintain
competes with the substrate for
.

D. As temperature increases beyond the

more enzyme * rate otrxn .

binding at the
Enlymes do not provide the energy to overcome the Ed for rxns the specific conformation of the active site Rate otrxn inhibitor
'

required to break down substrates is limited by enzyme

rxns in water is
Catalyse CMV me active site The
frequency
.

optimum temperature the rate of rxn are disrupted cone of enzyme -

.

in loss of 3D conformation
.

Energy resulting
, .

carbohydrates
Hydrolases into simpler substances eg lipases proteases decreases despite molecules having 4 > ENZYME conc substrate collisions is similar to the frequency of
,
.
, .

otenlyme & its active site The enzyme is

.

- Kinetic energy Heat has disrupted the denatured As a result, the substrate can
.
.

no
thdte Of ✓ ✗ No enzyme inhibitor collisions
-
. .
:
, the no Ot .
E -
s
.

V10 longer bind to the active site of the enzyme N complexes formed is abt the same as E- 1
catalyse the joining of two molecules ( ligation > by forming enzyme hydrogen bonds and hydrophobic ,

rate of formation of E- s complexes decreases, Rate of unlimited complexes formed & this leads to allow rate of rxn
ligases interactions within the secondary h by /
-

ligase involved in DNA

,
DNA
a new chemical bond , eg
-

a- hence decreasing the rate of formation of

axrdteotvxhenzym.ec#,
products
replication .

EAW Activation energy tertiary structures of the enzymes ,

v. enzyme
v10 enzyme .

of specific 3D ✓ may
resulting in the loss
.

enzyme At high substrate conc the substrate can outcompete

'

other than
Cleave various bonds by means hydrolysis
,
y
Lyases conformation of emyme hits active site
{ oxidation
.

the competitive inhibitors for binding to the active

site of the entry me , so the frequency of enzyme substrate
.

The emymc is denatured The substrate

-

2 eactdnts .

,
,
-

can no longer bind to the active site of the collisions is higher than the frequency of enlyme inhibitor -

Overall E S complexes , t the rdtc

catalyse interconversion of isomers energy enzyme to form E- 1
-

Isomerases the collisions : , more E -

s complexes than complexes are
of formation of 't s complexes hence t
.

released during
.

produce, rxn
I formed , leading rate otrxh
of higher
-

to a
.
, .

the rate of formation of products .

- Rate rxn limited by Km

> Time substrate cone •

At very high substrate conc , substrates can

out
-

Tvncnanged

/
,

Rate of rxn CK -1101°C For binding -10

compete the competitive inhibitors
① ,, =
Rate of rxndtxic 1 rate
the active site of the enzyme { allow the Mdx
.

1
Enzyme reduce the activation of rxn Climax ] to be reached
'

energy by either
.

(d) holding the substrates close together at the correct angle and orientation at active site for I Substrate cone
-
Thus , competitive inhibition is
&
.

c- A ← B → reversible can be overcome by high

•
-
→
successful lnleractron and collision-

( anabolic tbh ) subs tra I e -

concentration .

(b) Straining the chemical bonds within the substrates until break ( catabolic rxnt
they .

Ratenotrxnlmoldm -3s
- i

inhibitors
3. Enzyme specificity
VMDX
-11 IVO
\
.

Enzymes are highly specific In the rxns they catalyse The generally catalyse the transformation of
. one
I
competitive inhibitors .

of restricted grp of similar substrate molecules Allosteric

particular type substrate molecule or a
Enzymes
I
.

t Vmax
Enzymes are
highly specific because : only substrates of complementary shape will { bind to an active ,
1
-

enter
site with specific 3D conformation spatial fit -

'

Allosteric enzymes are enzymes that exist in two other forms active { / I
Enzyme {
inactive
-

i substrate must be chemically

compatible , i. e have charge and
.

hydrophilic hydrophobic complementary Chemical fit

- -

Most allosteric ally regulated eniymes are made up of two or more polypeptide chains ( subunits ) each w its own
1 .
,
1
active site Allie steric sites are located where subunits are joined .

I
.

Regulatory compounds allosteric activators and allosteric inhibitorsbind to the allosteric enzyme at its allosteric
y
- -

1 > Substrate concentration

Chemical fit site , a specific site located away from the active site , resulting in the reversible change in the structure of the
oscillate btw the active { inactive
-3
and allowing the enzyme to km.no < km inhibitor IMO / dm
enlyme ,
active site conformation .

inhibitor
Besides having spatial fit for an enzyme -

substrate complex to
•

The
binding
otdndlloesterio activator to the allosteric site converts the allosteric enzyme from inactive '
For all substrate conc UP to K , the rate of rxn

lock { Key hypothesis Induced fit -

hypothesis form the enzyme & substrate Csi must

, also be chemically compatible to active form .
Is aways lower compared when there are no

& capable of bonding u each other inhibitors

The Initial Shape Of the active site of the enzyme
The active form of the enzyme is shaped
.

may not be exactly such that the substrate has a complementary shape to that of
•

'
'
The substrate can be visualised
-

as the key shape

Enzyme & substrate ( s ) are usually held tgt temporarily by weak
whose When substrate conc
• •

it v. high ( i. e right
complementary to that of the substrate V1
& & bind site form
'

the active site the substrate can enter to the active to E S complex
is
-

side of 4) the rate of rxn is the same

complementary bonds leg ionic bonds hydrogen bonds hydrophobic interactions )
.

to that of the enzyme 's active site , , ,

substrate enters h binds to the active site , it induces binding of not 11 is present )
. ,

whether
,
lock However The an allosteric inhibitor to the allosteric site converts the allosteric site converts the allosteric or
' '
'
as the
-

-
the that be formed h broken readily during the transition
.

can
.
,

'

When an enzyme -

substrate complex is formed ,

chances
d conformational change In the shape of the enzyme such that the state . enzyme from active to inactive form .

of successful rxn increase .

Substrate can fit even more
snugly into the active site , allowing '

The type of substrate that enters and binds to the active site of
Summary of the differences btw competitive
& non - competitive inhibitors

enhanced interaction b/w the chemical groups of the substrate { the the
enzyme depends on the nature of amino acids making up
-

The inactive form of enzyme is shaped such that the substrate does not have a
complementary shape to that of the active
Once pelts are formed Camino acid ) residues at the active site for successful rxn hence
binding
'

longer fit into the

can occur
, they no active catalytic . Its active site .
site ,
no .

Competitive inhibitor Non -

Competitive inhibitor
site and are released into the surrounding medium
-

If the exposed The areas of contact between the subunits of an allosteric enzyme fittest in such
. 12 groups of these amino acids are
-

a way that a conformational

leaving the active site free to receive new substrate molecules reactive portion
electrically charged , then the of
change in one subunit is transmitted to all others Thus a single activator or inhibitor that binds -10 One Structurally similar to the Not structurally
.

.
, molecule substrate
similar to the substrate .

the substrate must also be polar or allosteric site will affect the active sites of all subunits
charged
complementarily .

✓ Allosteric activators / Inhibitors

& Binds It
.

Enters binds Site other than ( allosteric site )

fluctuating
The to the active site d the active site
activity of an allosteric emyme changes in response to conc of the compounds In
-

0 A non polar substrate cannot react with the active

- .

regulatory .

cofactors site of such an enzyme even if it does , by chance fit this

,

way , allosteric enzymes control the rates of key rxns in metabolic pathways .

into the active site Binds

.

Birds temporarily to enzyme at active site temporarily or

permanently to en Lyme at duo esoteric site

Cofactors are non fn etticientiy There are three types of cofactors prosthetic grps { coenzymes Doesnot compete with substrate for active site but changes the
protein components that are essential for some enzymes to Inorganic ions End Product Inhibition
-
-
.
. ,
for { prevent , formation of
conformation of the active site , thus preventing formation of E- s
compete, w substrate +ne active site
1. Inorganic Ions E- s complex ,
complex .

small metal ions

Usually
'
.

Metabolic Rate of reaches Max C similar normal rxn in the Rate of reaches d lower maximum level Cds compared to norm "
non
pathways rxn level
the formation
to
They assist in
by moulding the Ehly me Into
.

of the enzyme
'
-
substrate complex a more suitable shape
.

substrate conc )
inniyi.gr , buy higher , in the absence of inhibitors even at a higher
.

of ,, a gun , + rate con , , , , we , ,

.

eg Calcium ( Ca 2-17 thrombokinase which into thrombin

Many
absence rxn
,

ions activate then prothrombin during blood

involve
.

converts clotting metabolic processes a series of enzyme catalysed rxns in which the put from the substrate for
- -

. .
one rxn acts as
the next rxn .

Effect of Inhibition can be over conned by adding Effect of inhibition cannot be corned by adding
{ this
over
Edlhrxn Is catalysed by different more
-

a allows intermediates to go down diff pathways more substrate

enzyme Substrate
.

Prosthetic
.

2. groups .

The different enzymes that catalyse Often form a linear series bound to membranes within the cell
-

such chain rxns

>
These are large organic molecules covalently { permanently bound to the en Lyme
,
Degree of inhibition depends the relative cone at
inhibition does not depend on the relative conc Of inhibitor
Degree
on
enzyme complex , eg pyruvate dehydrogenase complex is a complex of three enzymes that of
constituting which
, .

a multi
.

the iron containing prosthetic group of cytochrome c oxidase transfers oxygen in the transport
-

eg Hdem is -

which electrons to electron chain

.

acetyl CoA by a process called pyruvate decarboxylation during &

.

inhibitor substrate
.

aerobic respiration
pyruvate into
,
Harem in catalase converts -

and substrate
eg
- . .

.
.

Such close proximity btw enzymes is efficient since collisions enzymes h their substrates are made more likely
•

btw .

No ettect on the km value 1

Enzyme 's affinity for substrate
3- Coenzymes km value is T / Enzyme 's
affinity for substrate ¥
End product inhibition remains the same .

-
an example of allosteric inhibition
These are small substances which unlike prosthetic grips are not bound to enzymes
.

organic permanently For metabolic a series of enzyme the final Pdf Of the pathway is usually
-

many pathway involving catalysed rxns an

, •
, , . -
,

It is
only during that these molecules loosely { allosteric inhibitor of one of the pathway earlier enzymes in the
-

rxns are non covalently associated with enzymes

- .
.

coenzymes generally derived from Such inhibition of an earlier

stage in a process by the final pdf is termed re feedback inhibition
' '

are vitamins .
-

In this way , an accumulation of the final Pdt will thus slow down or stop its further product -^ preventing wastage ,
of
resources .

Zymogen s -
Inactive precursors of enzymes -

When the product is used up , the innit ion is lifted & production is switched back on again .

Some enzymes are sytnesized in the inactive forms known

Being in nature negative feedback inhibition is thus
-

Zymogen self regulatory important in the metabolism of

as or pro coordinating
•

enzymes
- -

.
,

These forms 4 cells preventing both shortage & overproduction

properly aligned to form
-

are inactive because although the catalytic contact residues are present , they are not the active site .
, of end products .

'
certain peptide bonds within the zymogen need to be cleared so that a new 3D conformation can be achieved { the active form of enzyme released .

eg Pepsin
.
is expressed as a pro form zymogen
-
i. e. pepsinogen , whose
primary structure has an additional 44 amino acids .