1. Bagimana percobaan Henry bisa mendapatkan pers (3.

3)
The major contribution of Henri was to think of enzyme reactions in two stages. In the first, the
substrate binds reversibly to the enzyme, forming the enzyme-substrate complex. This is
sometimes called the Michaelis complex. The enzyme then catalyzes the chemical step in the
reaction and releases the product.
To find the maximum speed of an enzymatic reaction, the substrate concentration is increased
until a constant rate of product formation is seen. This is shown in the saturation curve on the
right. Saturation happens because, as substrate concentration increases, more and more of the
free enzyme is converted into the substrate-bound ES form. At the maximum velocity (Vmax) of
the enzyme, all the enzyme active sites are bound to substrate, and the amount of ES complex is
the same as the total amount of enzyme. However, Vmax is only one kinetic constant of enzymes.
The amount of substrate needed to achieve a given rate of reaction is also important. This is
given by the Michaelis-Menten constant (Km), which is the substrate concentration required for
an enzyme to reach one-half its maximum velocity.
Each enzyme has a characteristic Km for a given substrate, and this can show how tight the
binding of the substrate is to the enzyme. Another useful constant is kcat, which is the number of
substrate molecules handled by one active site per second.
2. Jelaskan mengapa ada fenomena temperature dan pH naik, aktifitas enzim juga naik, sampai
mencapai temperature/pH optimum, aktifitas enzim menurun
The effect of temperature on the rate of enzyme-controlled reactions
A typical graph of rate against temperature might look like this:
The temperature at which the rate is fastest is called the optimum temperature for that enzyme.
Higher temperature generally causes more collisions among the molecules and therefore
increases the rate of a reaction. More collisions increase the likelihood that substrate will collide
with the active site of the enzyme, thus increasing the rate of an enzyme-catalyzed reaction.
Above a certain temperature, activity begins to decline because the enzyme begins to denature.
Above this temperature the enzyme structure begins to break down (denature) since at higher
temperatures intra- and intermolecular bonds are broken as the enzyme molecules gain even
more kinetic energy.
The rate of chemical reactions therefore increases with temperature but then decreases as
enzymes denature.
Different enzymes have different optimum temperatures. Some enzymes, for example, in
organisms known as thermophiles or extremophiles are capable of working at temperatures like
80C or even higher.
The optimum temperature for a particular enzyme varies depending on how long it is exposed to
the higher temperatures. Enzymes can withstand temperatures higher than their normal optimum
if they are only exposed to the higher temperatures for a very short time.
A reminder about the effect of temperature on rate in ordinary chemical reactions
Remember that for molecules to react, they have to collide with an energy equal to or greater
than the activation energy for the reaction.
Heating a reaction makes the molecules move faster and so collide more often. More collisions
in a given time means a faster reaction.
But far more importantly, increasing the temperature has a very big effect on the number
of collisions with enough energy to react. Quite a small temperature rise can produce a
large increase in rate.

As a reasonable approximation for many (although not all) reactions close to room temperature,
a 10C increase in temperature will double the rate of a chemical reaction. This is only an
approximation - it may take 9C or 11C or whatever, but it gives you an idea of what to expect.
Explaining the rate against temperature graph
At lower temperatures, the shape of the graph is exactly what you would expect for any
reaction. All that needs explaining is why the rate falls above the optimum temperature.
Remember that the enzyme works because its substrate fits into the active site on the protein
molecule. That active site is produced because of the way the protein is folded into its tertiary
structure.
Heating an enzyme gives the protein chains extra energy and makes them move more. If they
move enough, then the bonds holding the tertiary structure in place will come under increasing
strain.
The weaker bonds will break first - van der Waals attractions between side groups, and then
hydrogen bonds. As soon as these bonds holding the tertiary structure together are broken, then
the shape of the active site is likely to be lost.
Obviously, the longer the enzyme is held at the higher temperature, the more time there is for
the enzyme structure to be broken up. Given enough time and high enough temperatures, the
enzyme structure is broken permanently. The enzyme is said to be denatured. For example,
boiling an egg for 5 minutes denatures the proteins in the egg. Once that has happened, you can't
un-boil it by cooling it!
The effect of pH on the rate of enzyme-controlled reactions
Each enzyme has an optimal pH.
A change in pH can alter the ionization of the R groups of the amino acids. When the charges on
the amino acids change, hydrogen bonding within the protein molecule change and the molecule
changes shape. The new shape may not be effective.
In the same way that every enzyme has an optimum temperature, so each enzyme also has an
optimum pH at which it works best.
For example, trypsin and pepsin are both enzymes in the digestive system which break protein
chains in the food into smaller bits - either into smaller peptide chains or into individual amino
acids.
Pepsin works in the highly acidic conditions of the stomach. It has an optimum pH of about 1.5.
On the other hand, trypsin works in the small intestine, parts of which have a pH of around 7.5.
Trypsin's optimum pH is about 8.
If you think about the structure of an enzyme molecule, and the sorts of bonds that it may form with
its substrate, it isn't surprising that pH should matter.
Suppose an enzyme has an optimum pH around 7. Imagine that at a pH of around 7, a substrate
attaches itself to the enzyme via two ionic bonds. In the diagram below, the groups allowing ionic
bonding are caused by the transfer of a hydrogen ion from a -COOH group in the side chain of one
amino acid residue to an -NH2 group in the side chain of another.
In this simplified example, that is equally true in both the substrate and the enzyme.

Now think about what happens at a lower pH - in other words under acidic conditions.
It won't affect the -NH3+ group, but the -COO- will pick up a hydrogen ion.
What you will have will be this:
You no longer have the ability to form ionic bonds between the substrate and the enzyme. If those
bonds were necessary to attach the substrate and activate it in some way, then at this lower pH, the
enzyme won't work.
What if you have a pH higher than 7 - in other words under alkaline conditions.
This time, the -COO- group won't be affected, but the -NH3+ group will lose a hydrogen ion.
That leaves . . .
Again, there is no possibility of forming ionic bonds, and so the enzyme probably won't work this
time either.
3. Jelaskan cofactor dan peranannya dalam enzymatic
Cofactors & Coenzime
Many enzymes require the presence of an additional, nonprotein, cofactor.
Some of these are metal ions such as Zn2+ (the cofactor for carbonic anhydrase), Cu2+, Mn2+,
K+, and Na+.
Some cofactors are small organic molecules called coenzymes. The B vitamins
o thiamine (B1)
o riboflavin (B2) and
o nicotinamide are precursors of coenzymes.
Coenzymes may be covalently bound to the protein part (called the apoenzyme) of enzymes as a
prosthetic group. Others bind more loosely and, in fact, may bind only transiently to the enzyme as
it performs its catalytic act.
Some enzymes do not need any additional components to show full activity. However, others
require non-protein molecules called cofactors to be bound for activity. Cofactors can be either
inorganic (e.g., metal ions and iron-sulfur clusters) or organic compounds (e.g., flavin and heme).
Organic cofactors can be either prosthetic groups, which are tightly bound to an enzyme, or
coenzymes, which are released from the enzyme's active site during the reaction. Coenzymes
include NADH, NADPH and adenosine triphosphate. These molecules transfer chemical groups
between enzymes.[44]
An example of an enzyme that contains a cofactor is carbonic anhydrase, and is shown in the ribbon
diagram above with a zinc cofactor bound as part of its active site.[45] These tightly bound molecules
are usually found in the active site and are involved in catalysis. For example, flavin and heme
cofactors are often involved in redox reactions.
Enzymes that require a cofactor but do not have one bound are called apoenzymes or apoproteins.
An apoenzyme together with its cofactor(s) is called a holoenzyme (this is the active form). Most
cofactors are not covalently attached to an enzyme, but are very tightly bound. However, organic
prosthetic groups can be covalently bound (e.g., thiamine pyrophosphate in the enzyme pyruvate
dehydrogenase). The term "holoenzyme" can also be applied to enzymes that contain multiple
protein subunits, such as the DNA polymerases; here the holoenzyme is the complete complex
containing all the subunits needed for activity.

Coenzymes
Space-filling model of the coenzyme NADH
Coenzymes are small organic molecules that transport chemical groups from one enzyme to
another. Some of these chemicals such as riboflavin, thiamine and folic acid are vitamins
(compounds which cannot be synthesized by the body and must be acquired from the diet). The
chemical groups carried include the hydride ion (H-) carried by NAD or NADP+, the acetyl group
carried by coenzyme A, formyl, methenyl or methyl groups carried by folic acid and the methyl
group carried by S-adenosylmethionine.
Since coenzymes are chemically changed as a consequence of enzyme action, it is useful to
consider coenzymes to be a special class of substrates, or second substrates, which are common to
many different enzymes. For example, about 700 enzymes are known to use the coenzyme NADH.
[47]

Coenzymes are usually regenerated and their concentrations maintained at a steady level inside the
cell: for example, NADPH is regenerated through the pentose phosphate pathway and Sadenosylmethionine by methionine adenosyltransferase
4. Jelaskan yang dimaksud dengan competitive inhibition, non inhibition, partially
competitive, juga mekanisme dan artinya
Inhibition
Competitive inhibitors bind reversibly to the enzyme, preventing the binding of substrate. On the
other hand, binding of substrate prevents binding of the inhibitor. Substrate and inhibitor compete
for the enzyme.
Types of inhibition. This classification was introduced by W.W. Cleland.
Enzyme reaction rates can be decreased by various types of enzyme inhibitors.
Kinetic scheme for reversible enzyme inhibitors.
Enzyme inhibitors are molecules that reduce or abolish enzyme activity. These are either reversible
(i.e., removal of the inhibitor restores enzyme activity) or irreversible (i.e., the inhibitor
permanently inactivates the enzyme).
Reversible inhibitors
Reversible enzyme inhibitors can be classified as competitive, uncompetitive, non-competitive or
mixed, according to their effects on Km and Vmax. These different effects result from the inhibitor
binding to the enzyme E, to the enzymesubstrate complex ES, or to both, as shown in the figure to
the right and the table below. The particular type of an inhibitor can be discerned by studying the
enzyme kinetics as a function of the inhibitor concentration. The four types of inhibition produce
LineweaverBurke and EadieHofstee plots that vary in distinctive ways with inhibitor
concentration. For brevity, two symbols are used:

and

where Ki and K'i are the dissociation constants for binding to the enzyme and to the enzyme
substrate complex, respectively. In the presence of the reversible inhibitor, the enzyme's apparent
Km and Vmax become (/')Km and (1/')Vmax, respectively, as shown below for common cases.

Ki only (

Type of
inhibition
competitive

Ki' only (

uncompetitive

Ki = Ki' (

) non-competitive

Ki Ki ' (

Km apparent

Vmax apparent

mixed

Non-linear regression fits of the enzyme kinetics data to the rate equations above can yield accurate
estimates of the dissociation constants Ki and K'i.
Irreversible inhibitors
Enzyme inhibitors can also irreversibly inactivate enzymes, usually by covalently modifying active
site residues. These reactions, which may be called suicide substrates, follow exponential decay
functions and are usually saturable. Below saturation, they follow first order kinetics with respect to
inhibitor.
Irreversible inhibitors react with the enzyme and form a covalent adduct with the protein. The
inactivation is irreversible. These compounds include eflornithine a drug used to treat the parasitic
disease sleeping sickness. Penicillin and Aspirin also act in this manner. With these drugs, the
compound is bound in the active site and the enzyme then converts the inhibitor into an activated
form that reacts irreversibly with one or more amino acid residues.
Competitive inhibition
In competitive inhibition, the inhibitor and substrate compete for the enzyme (i.e., they can not bind
at the same time). Often competitive inhibitors strongly resemble the real substrate of the enzyme.
For example, methotrexate is a competitive inhibitor of the enzyme dihydrofolate reductase, which
catalyzes the reduction of dihydrofolate to tetrahydrofolate. The similarity between the structures of
folic acid and this drug are shown in the figure to the right bottom. Note that binding of the
inhibitor need not be to the substrate binding site (as frequently stated), if binding of the inhibitor
changes the conformation of the enzyme to prevent substrate binding and vice versa. In competitive
inhibition the maximal velocity of the reaction is not changed, but higher substrate concentrations
are required to reach a given velocity, increasing the apparent Km.
Uncompetitive inhibition
In uncompetitive inhibition the inhibitor can not bind to the free enzyme, but only to the EScomplex. The EIS-complex thus formed is enzymatically inactive. This type of inhibition is rare,
but may occur in multimeric enzymes.
Non-competitive inhibition
Non-competitive inhibitors can bind to the enzyme at the same time as the substrate, i.e. they never
bind to the active site. Both the EI and EIS complexes are enzymatically inactive. Because the
inhibitor can not be driven from the enzyme by higher substrate concentration (in contrast to
competitive inhibition), the apparent Vmax changes. But because the substrate can still bind to the
enzyme, the Km stays the same.
Mixed inhibition

This type of inhibition resembles the non-competitive, except that the EIS-complex has residual
enzymatic activity.
In many organisms inhibitors may act as part of a feedback mechanism. If an enzyme produces too
much of one substance in the organism, that substance may act as an inhibitor for the enzyme at the
beginning of the pathway that produces it, causing production of the substance to slow down or stop
when there is sufficient amount. This is a form of negative feedback. Enzymes which are subject to
this form of regulation are often multimeric and have allosteric binding sites for regulatory
substances. Their substrate/velocity plots are not hyperbolar, but sigmoidal (S-shaped).
The coenzyme folic acid (left) and the anti-cancer drug methotrexate (right) are very similar in
structure. As a result, methotrexate is a competitive inhibitor of many enzymes that use folates.
Uses of inhibitors
Since inhibitors modulate the function of enzymes they are often used as drugs. An common
example of an inhibitor that is used as a drug is aspirin, which inhibits the COX-1 and COX-2
enzymes that produce the inflammation messenger prostaglandin, thus suppressing pain and
inflammation. However, other enzyme inhibitors are poisons. For example, the poison cyanide is an
irreversible enzyme inhibitor that combines with the copper and iron in the active site of the enzyme
cytochrome c oxidase and blocks cellular respiration.
5. Apa perbedaan activitas enzim dengan stabilitas enzim
Control of activity
There are five main ways that enzyme activity is controlled in the cell.
1. Enzyme production (transcription and translation of enzyme genes) can be enhanced or
diminished by a cell in response to changes in the cell's environment. This form of gene
regulation is called enzyme induction and inhibition (see enzyme induction). For example,
bacteria may become resistant to antibiotics such as penicillin because enzymes called betalactamases are induced that hydrolyse the crucial beta-lactam ring within the penicillin
molecule. Another example are enzymes in the liver called cytochrome P450 oxidases,
which are important in drug metabolism. Induction or inhibition of these enzymes can cause
drug interactions.
2. Enzymes can be compartmentalized, with different metabolic pathways occurring in
different cellular compartments. For example, fatty acids are synthesized by one set of
enzymes in the cytosol, endoplasmic reticulum and the Golgi apparatus and used by a
different set of enzymes as a source of energy in the mitochondrion, through -oxidation.[68]
3. Enzymes can be regulated by inhibitors and activators. For example, the end product(s) of
a metabolic pathway are often inhibitors for one of the first enzymes of the pathway (usually
the first irreversible step, called committed step), thus regulating the amount of end product
made by the pathways. Such a regulatory mechanism is called a negative feedback
mechanism, because the amount of the end product produced is regulated by its own
concentration. Negative feedback mechanism can effectively adjust the rate of synthesis of
intermediate metabolites according to the demands of the cells. This helps allocate materials
and energy economically, and prevents the manufacture of excess end products. The control
of enzymatic action helps to maintain a stable internal environment in living organisms.
4. Enzymes can be regulated through post-translational modification. This can include
phosphorylation, myristoylation and glycosylation. For example, in the response to insulin,
the phosphorylation of multiple enzymes, including glycogen synthase, helps control the

synthesis or degradation of glycogen and allows the cell to respond to changes in blood
sugar.[69] Another example of post-translational modification is the cleavage of the
polypeptide chain. Chymotrypsin, a digestive protease, is produced in inactive form as
chymotrypsinogen in the pancreas and transported in this form to the stomach where it is
activated. This stops the enzyme from digesting the pancreas or other tissues before it enters
the gut. This type of inactive precursor to an enzyme is known as a zymogen.
5. Some enzymes may become activated when localized to a different environment (eg.
from a reducing (cytoplasm) to an oxidising (periplasm) environment, high pH to low pH
etc). For example, hemagglutinin in the influenza virus is activated by a conformational
change caused by the acidic conditions, these occur when it is taken up inside its host cell
and enters the lysosome.
17.a Mengapa produk degradasi lignin susah dimanfaatkan ?
Kayu umumnya terdiri dari beberapa senyawa complex yaitu -selulosa, hemiselulosa, lignin
dan glycoprotein extension. Kandungan lignin dalam kayu sekitar 25% berat kering. Lignin
adalah polimer dari alcohol aromatic yang cukup kompleks dan besar sehingga sulit untuk
dimanfaatkan selain untuk bahan bakar
17.b Mengapa produk degradasi hemiselulosa lebih mudah dimanfaatkan ?
Hemiselulosa dengan beberapa variasi yang spesifik yang berbeda untuk tiap tanaman,
merupakan cabang polimer dari pentose (xilose dan arabinose) dan beberapa hexose
(glucose,galactose dan mannose) , karena po;omer tergolong pendek dan lebih sederhana
maka produk degradasi hemisoluluse bisa banyak dimanfaatkan, antara lain sebagai bahan
baku furfural.
17c. Jenis enzyme dalam degradasi selulosa dam hemiselulosa
Application
Paper industry

Enzymes used
Amylases,
Xylanases,
Cellulases
Ligninases
Cellulases

Biofuel industry

Uses
Degrade starch to lower viscosity, aiding sizing
and coating paper. Xylanases reduce bleach
required for decolorising; cellulases smooth fibers,
enhance water drainage, and promote ink removal;
lipases reduce pitch and lignin-degrading enzymes
remove lignin to soften paper.
Used to break down cellulose into sugars that can
be fermented (cellulosic ethanol).

Cellulose in 3D
Ligninase
Enzim Hemiselulose

Use of lignin waste

Endo-1.4--D-xylanase
Endo-1.4--D-mannanase
Endo-1.4--D-xylosidase
Endo-1.4--D-mannosidase
Endo-1.4--D-glcosidase
-L-arabinosidase
-D-glucuronidase
-D-galactosidase
Acetyl xylan esterase
Acetyl galactoglucomanan
esterase

Depolimerisasi rantai utama

hemiselulosa
Hidrolisa pada gugus
oligosakarida
Pemutusan gugus disisi samping
hemiselulose
Pemutusan grup pada sisi yang
teresterifikasi

Enzim Selulose

Endo--1.4-gluconase

Menyerang rantai bagian dalam

dari selulosa amorphous
menghasilkan selodextrin,
sellobiosa atau glucose

Exo--1.4-gluconase

Menyerang crystalline cellulose

-1.4-glucosidase

Menghidrolisa seloboisa
menjadi glucose dan degradasi
seloligosacharida

23. Spesific activity : U/mg protein, merupakan unit activity dari protein, parameter ini dibutuhkan
dalam menentukan konsentrasi enzim untuk substrat tertentu pada temperatur dan waktu reaksi
tertentu terkait dengan aktifitas enzimnya. Unit of actifity (U) tiap enzim berbeda.
24. a.Mengapa enzim perlu diimobilisasi ?
Enzim adalah protein yang mudah larut dalam air, sehingga sangat sulit untuk memisahkan
enzim dalam rangka untuk digunakan kembali (reused). Enzim dapat diimobilisasi pada
permukaan atau dalam insoluble matrix dengan cara kimia atau fisik
b.Jelaskan teknik Immobilisasi Enzim ?
25. Apa yang dimaksud dengan external dan internal mass transfer resistance.
- External Mass-Transfer Resistance : Immobilisasi enzim pada permukaan insoluble particel
- Internal Mass-Transfer Resistance : Immobilisasi enzim dengan copolymerization atau
microencap sulation
26. Mengapa enzim yang diimmobilisasi memberikan fenomena yang tdk tergantung pada MichMenten ?
Pada enzim yang diimobilisasi, laju reaksi melibatkan dua hal yaitu : perpindahan massa dan
kinetika reaksi enzim, laju ini dinyatakan sebagai overall rate reaction.
Untuk itu perlu diketahui laju yang mengontrol terlebih dahulu.