METABOLISM

and
ENZYME
Alfi Muntafiah
Laboratorium Biokimia FK Unsoed
Metabolism is all chemical reactions that occur in the body

 Two types of metabolism

 Anabolism :
provides the substances needed for cellular growth and repair
• larger molecules are made
• requires energy
 Catabolism :
breaks down larger molecules into smaller ones
• larger molecules are broken down
• releases energy

Both breakdown and buildup reactions have activation energies

Breakdown Reactions Release Energy: Exergonic/exothermic

Energy Level
Z Activation
X + Y + Energy
Z
X+Y

Time

Buildup Reactions Absorb or Require Energy: Endergonic/endothermic

A + B C
+ ATP

Energy Level
C Activation
Energy
A+ B

Time
A metabolic pathway is the sequence of biochemical
reactions in order to transfer chemical energy

Enzyme 1 Enzyme 2 Enzyme 3

A B C D
Reaction 1 Reaction 2 Reaction 3
Starting Product
molecule

It is begins with a specific molecule (reactant) and ends with a product

Each step of biochemical reaction is catalyzed by a specific enzyme
In the laboratory,
we heat the reactants in
order to provide activation
energy for a chemical
reaction
Inside the cell,
a different mechanism is
required as heating up the
reactants is not possible
Enzymes are giant macromolecules
which catalyse biochemical reactions.

 They are increase the rate of reaction but they themselves

are not changed by the reaction.
 Each enzyme catalyses a single chemical reaction on a
particular chemical substrate
 Enzyme can work inside cells and outside cells
 Their three-dimensional structures are highly complex, yet they
are by spontaneous folding of a linear polypeptide chain.
 Sebagian besar dihasilkan oleh sel hidup, dapat diekstraksi
dari sel, tanpa kehilangan aktifitas biologiknya.
Enzymes Are Biological Catalysts

Figure 5.2
 Enzymes control rates of metabolic reactions

• lower activation energy needed

to start reactions
• substrate specific
• shape of active site
determines substrate
 Enzyme names reflect the  Faktor-faktor yang mempengaruhi
substrate reaksi enzimatis :
 Kadar enzim
• have the suffix – ase
 Kadar substrat
• example : sucrase, lactase,
 Suhu
protease, lipase
 pH
 Koenzim
 inhibitor
 Bila substrat dinaikkan kadarnya, maka kecepatan reaksi juga
akan meningkat. Peningkatan reaksi sesuai dengan
peningkatan kadar substrat sampai enzim dijenuhi oleh
substrat.
 Kecepatan awal reaksi berbanding lurus dengan kadar enzim
 Pengaruh kadar koenzim terhadap kecepatan reaksi =
pengaruh enzim terhadap kecepatan reaksi
 Enzymes can be denatured by temperature and pH

Pada pH rendah atau tinggi, enzim

mengalami denaturasi, sehingga
aktifitas enzim menurun
Perubahan pH yang tinggi
menyebabkan perubahan
konformasi enzim sehingga enzim
menjadi tidak aktif.

Suhu optimal merupakan suhu

dimana reaksi berjalan dengan
cepat. Bila suhu melebihi suhu
optimal, maka reaksi akan menurun
karena enzim mengalami denaturasi.

Figure 5.6
An enzyme inhibitor is a substance that inhibits the
catalytic action of the enzyme.
 Competitive inhibition

Figure 5.7a, b
An enzyme inhibitor is a substance that inhibits the
catalytic action of the enzyme.

 Noncompetitive inhibition

ATP, pyruvate, end

amino acid
Figure 5.7a, c
 Enzymes catalyze the thousands of reactions that
need to take place in order to maintain life

What are some of these reactions?

 digestion
 respiration
 photosynthesis
(plants and some bacteria)
 protein synthesis.
DIGESTIVE
ENZYMES

Alfi Muntafiah
Laboratorium Biokimia
FK UNSOED
Digestion is the process of breaking down food

 Nutrients in food must first be broken down into particles

that are small enough to be transported and absorbed by
the epithelium of the small and large intestine,
Digestion occurs through two different
processes :

 physical digestion,
large chunks of food are ground into tiny particles
 chemical digestion,
 through the use of enzymes released into the digestive tract
 large polymeric biomolecules are broken into individual
monomers or oligomers (e.g. dimers or trimers).
what we eat

glu
Chemical digestion is essential for breaking food into particles that can
be absorbed
glu
.

these are the molecules

that are small enough to
absorb into the
bloodstream (not drawn
to scale)
Digestion occurs primarily within digestive tract =
GI (gastrointestinal) tract = alimentary canal
 There are many different substances that are secreted into the
different segments of the digestive tract :
 Mucus,
 bile salts,
 bilirubin,
 hydrochloric acid (HCl), and
 sodium bicarbonate (NaHCO3)
They are just some of the substances mixed with the food as it
passes through the digestive tract, and many of these substances
facilitate the breakdown of food.

 However, the most important substances secreted for the

purpose of digestion are the digestive enzymes.
digestive enzyme are produced and secreted by the
cells from almost all parts of the digestive system

 Salivary glands
 Lingual glands
 Stomach
 Pancreas
 Liver
 Intestinal mucosa

Often final steps of digestion

take place in the villi of
enterocytes
without the presence of these enzymes,chemical
digestion would essentially not occur.

 Digestive enzymes greatly enhance the rate at which the

covalent bonds that link subunits together to form polymeric
biomolecules are broken.

 Although substances such as HCl and NaHCO3 can alter

noncovalent bonding patterns within and among
biomolecules, they typically cannot break down covalent
bonds.
Not all enzymes work inside cells.
Digestive enzymes work outside cells.

These enzymes pass out of the cells

into mouth (saliva), the stomach
and small intestine.
Here the enzymes help to break
down large food molecules into
smaller molecules that are more
easily absorbed.
 Two factors that influence enzyme activity in the context
of digestion: temperature and pH.

 temperature can influence on the rate at which enzyme-catalysed

reactions proceed.
 Low temperatures result in slow reaction rates because overall kinetic
energy is reduced
 Very high temperatures can also slow the rate of an enzyme catalyzed
reaction because high temperatures destabilize the noncovalent
interactions in the enziyme.
 non covalent interaction give enzymes the specific tertiary and
quaternary structures that enable them to function as catalysts.

 This highlights the importance of the stable core body temperatures

of humans in their digestive processes.
 Two factors that influence enzyme activity in the context
of digestion: temperature and pH.

 Enzyme activity is also influenced by the

pH
of the surrounding fluid
 [H+] can induce changes in the tertiary
and quaternary structure of proteins.
 Thus enzymes have a particular pH where
they have the proper conformation to
have maximal catalytic activity,
 significant pH deviation will result in a
decrease in catalytic activity.
Enzymes Become Non-Functional at pH Extremes and High Temperatures

Stomach Enzyme within = folded, functional enzyme

enzyme a body cell = denatured, non-functional enzyme
Enzyme within
H Reaction
- rate is slow a body cell Enzyme from
+ H H OH- OH
+
+ at cold temperatures hot springs
(products formed per second)

(products formed per second)

H H because
OH - molecules bacterium
+ +
H encounter enzyme
Enzymatic rate

Enzymatic rate
+
+ OH- OH- less often
H OH-
+
H H OH-
+
+

H OH-
+ OH-
H H
+ +
H OH-
+

0 2 4 6 8 10 12 10 20 30 40 50 60 70
pH (in pH units) Temperature (oC)
 pH varies widely among different segments of
the digestive tract

 It will effectively restricting the function of that enzyme to a

particular region of the digestive tract

 Altering pH among different regions of the digestive tract

effectively enables a stepwise process of chemical
digestion, where enzymes are activated for digestion at
one point and then deactivated at the next.
 different enzymes have maximum
catalytic activity at these various pH’s

 gastric enzymes such as pepsin have

maximum catalytic activity at the
very low pH of the stomach, and no
longer function once moved into
the alkaline conditions of the small
intestine.
 intestinal and pancreatic enzymes,
such as trypsin, function optimally at
moderately alkaline pH.
Carbohydrates are the first type of biomolecule
to be chemically digested in the digestive tract

chemical digestion begins in the oral cavity through a salivary

enzyme called salivary amylase (or ptyalin).
Salivary amylase begins the breakdown of the polysaccharide
amylose (starch, the principle storage carbohydrate in plants)
into the disaccharide maltose.
chemical digestion of carbohydrates effectively stops when
food is swallowed and transferred to the small intestine,.
Carbohydrates are the first type of biomolecule
to be chemically digested in the digestive tract

In the small intestine, the chyme is exposed to :

pancreatic amylase
which continues the process of breaking down starch and
glycogen into disaccharides and trisaccharides
brush-border enzymes
 lactase, sucrase, and maltase
break down specific oligosaccharides into the
monosaccharides that are absorbed by the intestinal
epithelium.
Chemical digestion of protein begins in the stomach.

 in response to neural stimulation (induced by smell, site and

taste of food), the lining of the stomach produces a mixture
of fluids called gastric juice
 as food enters  distension of the stomach
 pH changes induced as the more neutral pH food enters to
the acidic stomach
.
 Chemical digestion of protein begins in the
stomach.

 Gastric juice contains a number of substances, but the two most

important for initiating protein digestion are :
 hydrochoric acid (HCl)
the protease pepsin.
 HCl is secreted by parietal cells in the gastric mucosa.
 The low pH (~2) of the gastric juice aids protein digestion :
First, the low pH denatures the tertiary structures of ingested
protein, making them easier to digest enzymatically.
Secondly, the low pH is required for the activation of pepsin
 Pepsin is produced by chief cells of the gastric mucosa
in an inactive (zymogenic) form called pepsinogen.

 Pepsinogen is inactive
when released in the
gastric pits,
 but once it diffuses into
the lumen of the
stomach the acidic
conditions enable it to
have a weak proteolytic
activity.
 Pepsinogen can digest some ingested protein,
 more importantly pepsinogen molecules will partially
digest one another, and converting each other into the
fully active enzyme pepsin.
 Pepsin breaks peptide bonds between amino acids thus
it cleaves long polypeptides into shorter polypeptides.
 most of the digestion of protein takes place in
the small intestine.

Indeed, individuals with complete gastrectomies can

still completely digest protein
The proteases that function in the small intestine come
from two major sources:
membrane bound enzymes on the brush-border of
the intestinal mucosa
enzymes secreted from the pancreas.
Chemicals in the chime induce cells in the small
intestine to secrete hormones

 the hormones secretin,

stimulates water and bicarbonate secretion in the pancreas,
 The hormones cholecystokinin (CCK),
stimulates enzyme secretion in the pancreas.
 These hormones, in turn, cause the pancreas to release
pancreatic juice through the duodenal papilla, when food enters
the small intestine
 proteases are found within pancreatic juice, most are
released as zymogens (tripsinogen)

 Enzymes in the brush-border (enterokinase

(EN)) activate these zymogens 
converting it to its active form (trypsin).

 Trypsin, in turn digests other pancreatic

zymogens into their active forms.

 Tripsin digest the polypeptides into a

combination of free amino acids,
dipeptides, and tripeptides that are
absorbed by the intestinal epithelium.
Lipids (principally triglycerides, or fat) are a particularly
challenging group of biomolecules to digest chemically

 This is because fats, being hydrophobic, tend to aggregate in large

droplets within the fluid of the digestive tract
 minimizing the surface area of contact between the fat and the
surrounding water.
 Since the digestive enzymes are water soluble, they can only come into
contact with and digest those triglyceride molecules at the surface of
the droplets.
 Thus, for fat digestion to be efficient, these large droplets must be broken
into much smaller droplets and held in these smaller droplets in order to
increase surface area and enable fat-digesting enzymes (lipases)
adequate contact with their substrate.
 Although there is a gastric lipase secreted in the
stomach that causes a small amount of fat
digestion, almost all of the digestion of fat takes
place in the small intestine.

 Like protein digestion, fat digestion is achieved

through the activity of both pancreatic and
intestinal brush border lipases.
efficient fat digestion also involves the secretion of bile
from the liver and gall bladder.

 Bile contains a mixture of bilirubin, cholesterol, phospholipids, inorganic ions,

phospholipids, and bile salts.

 Secretion of bile into the small intestine is triggered by CCK (the same hormone
that induces pancreatic enzyme secretion).

 The bile salts are particularly important for fat digestion in that they serve as
emulsification agents.
 bile salts are amphipathic molecules

 Bile salt help to break the large fat droplets into

tiny emulsification droplets and prevent them
from reforming into large droplets

 This increases the surface area over which

lipases can come into contact with triglycerides
and hydrolyze them into : free fatty acids and
monoglycerides  absorbed by the intestinal
epithelium.
 Bile acid micelles
 Hydrophobic region of the bile
salt is oriented from the water
molecules x hydrophilic region
interacts with water

 Mixed micelles contain (beside

bile acids) phospholipids and
cholesterol, or FA and
acylglycerols; FA and
phospholipids form a bilayer in
the interior, bile salts occupy the
edge.
Not everything can be digested

 Many plant polymers, including celluloses, hemicelluloses, inulin,

pectin, are resistant to human digestive enzymes
 A small percentage of this „dietary fibre“ is hydrolyzed and then
anaerobically metabolized by the bacteria of the lower intestinal
tract
 This bacterial fermentation produces H2, CH4, CO2, H2S, acetate,
propionate, butyrate, lactate
enzymes

 enzymes cause chemical

reactions to occur
 they are extremely specific
 they only break down the
molecule they’re designed for
enzymes

 enzyme floats into place

enzymes
enzymes
enzymes

 it can only fit in one place

 for amylase -- only clips off END units
enzymes H2O

 water is needed also

enzymes H2O

 water floats into place

enzymes H2O

 breakage occurs
 enzymes enzyme unchanged

 free glucose ready for absorption

 enzyme ready for next reaction free glucose
 enzymes enzyme unchanged

 free glucose ready for absorption

 enzyme ready for next reaction free glucose
 enzymes enzyme unchanged

 free glucose ready for absorption

 enzyme ready for next reaction free glucose
 enzymes enzyme unchanged

 free glucose ready for absorption

 enzyme ready for next reaction free glucose
 enzymes enzyme unchanged

 free glucose ready for absorption

 enzyme ready for next reaction free glucose
 Vid.
terimakasih