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DIGESTIVE

ENZYMES

Bagian Biokimia
FK UNSOED
Enzymes

 giant macromolecules which catalyse biochemical reactions.


 Enzyme can work inside cells and outside cells
 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
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.
Digestion is the mechanical & chemical process of
breaking down food of 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)

one of the main


purposes of the
digestive system is to
turn food into these
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,
 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.
main digestive enzymes
 amylases
 break starch into monosaccharide units
 (clip off the free ends)

 lipases
 break the fatty acids off the glycerol
 proteases
 break down proteins from quaternary
structure down to individual amino acids
 thereare many different specific varieties
of each of these
digestive juices
1. saliva -- from salivary glands
 amylases
 water
2. bile -- made in liver, stored and released from gall
bladder
 emulsifier
 fat --> smaller GLOBS
 MECHANICAL breakdown of fats (NO change to chemical
structure of fats!)
3. pancreatic juice -- from pancreas
 baking soda, lipases, and other digestive enzymes
 CHEMICAL BREAKDOWN OF FATS
4. HCl -- stomach
 just HCl
5. digestive enzymes -- stomach
 mainly proteases
DIGESTION & ABSORBTION
OF CARBOHYDRATE
the first type of biomolecule to be chemically digested
begins in the oral cavity
through a salivary enzyme called salivary amylase (or
ptyalin).
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.
Carbohydrat digestion mostly occurs in small intestine
In the small intestine, the chyme is exposed to :
pancreatic amylase
brush-border enzymes (disaccharidases):
pancreatic amylase
continues the process of breaking down starch and glycogen
into disaccharides and trisaccharides
brush-border enzymes (disaccharidases):
 maltase breaks maltose into 2 glucoses
 Lactase breaks lactose into 1 glucose + 1 galactose
 Sucrase breaks sucrose into 1 glucose and 1 fructose
break down specific oligosaccharides into the
monosaccharides that are absorbed by the intestinal epithelium.
 transported to the liver & are converted to glycogen.
Mechanisms for Absorption of Monosaccharides :

 Glucose and galactose


are absorbed by a sodium-dependent process.
carried by transport protein (SGLT 1) and compete with each other for
intestinal absorption
 Other monosaccharides
are absorbed by carrier-mediated diffusion.
Because they are not actively transported, fructose and sugar alcohols are only
absorbed down their concentration gradient, and after a moderately high
intake, some may remain in the intestinal lumen, acting as a substrate for
bacterial fermentation.
Large intakes of fructose and sugar alcohols can lead to osmotic diarrhea.
Transport of glucose, fructose, and galactose
across the intestinal epithelium

 The SGLT 1 transporter is coupled to


the Na+-K+ pump, allowing glucose
and galactose to be transported against
their concentration gradients.
 The GLUT 5 Na+-independent
facilitative transporter allows
fructose, as well as glucose and
galactose, to be transported down their
concentration gradients.
 Exit from the cell for all sugars is via
the GLUT 2 facilitative transporter
DIGESTION & ABSORPTION
OF PROTEINS
Native proteins are resistant to digestion because
few peptide bonds are accessible to the proteolytic
enzymes without prior denaturation of dietary
proteins (by heat in cooking and by the action of
gastric acid).
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
.
Gastric juice

contains a number of substances, but the two most important for


initiating protein digestion are :
 hydrochoric acid (HCl)
 is secreted by parietal cells in the gastric mucosa
 The low pH (~2) of the gastric juice :
 denatures the tertiary structures of ingested protein, making
them easier to digest enzymatically.
 required for the activation of pepsin
 the protease pepsin.
Pepsin

produced by chief cells


 inactive (zymogenic)
form : pepsinogen.

Pepsinogen is inactive when


released in the gastric pits,
once it diffuses into the lumen of the stomach, the acidic conditions
enable it to have a weak proteolytic activity.
can digest some ingested protein
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.
  individuals with complete gastrectomies can still
completely digest protein
The major sources of proteases :
enzymes pancreas.
Trypsin &chymotrypsin break proteis into smaller and
smaller units.
Carboxypeptidase breaks peptides into free amino acids
membrane bound enzymes on the brush-border of the
intestinal mucosa
Chemicals in the chime induce cells in the small
intestine to secrete hormones :

hormones secretin
stimulates water and bicarbonate secretion in the pancreas,
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)
Prevents them from breaking down pancreas
(where they’re made &stored)
Activation occurs when they come in contact
with certain chemical found in the small
intestine.
Enzymes in the brush-border (enterokinase
(EN)) activate these zymogens  trypsin
Trypsin
digests other pancreatic zymogens into their active
forms.
digest polypeptides into a combination of free amino
acids, dipeptides, and tripeptides that are absorbed by
the intestinal epithelium.
DIGESTION & ABSORPTION
OF LIPIDS
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.
The major lipids in the diet are triacylglycerols and, to a
lesser extent, phospholipids
These are hydrophobic molecules
have to be hydrolyzed and emulsified to very small
droplets before they can be absorbed
Fat Digestion

1. Initiated by lingual and gastric lipase


2. Pancreatic lipases continues fat digestion
• subunits now cross into microvilli
• subunits are reassembled into triglycerides, combined with cholesterol,
and transported to the circulatory system.
Most fat digestion occurs in small intestine.

• Also involves the secretion of bile from the liver and gall
bladder
• The bile salts are particularly important for fat digestion in that
they serve as emulsification agents, so exposing more fat to
enzymes.
• Bile salts link fat molecules to water molecules; (normally fats
are hydrophobic).
bile salts

amphipathic molecules
help to break the large fat droplets into tiny emulsification
droplets
prevent them from reforming into large droplets
increases the surface area over which lipases come into contact
with triglycerides
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
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