Biokimia

Enzim
ENZIM
Suatu senyawa dengan kemampuan mengkatalis suatu
reaksi kimia melalui suatu mekanisme aktifasi yang
spesifik

Enzim: Protein

Bentuknya: Globular

Struktur 3-D »
Human pancreatic
amylase
 Enzim = Biokatalis
Rekasi berlangsung
pada kondisi fisiologis
pH 7.4, 37oC

Memproses jutaan
molekul tiap detiknya

Sangat Spesifik
Hanya mereaksikan
1 atau sedikit jenis
molekul
(substrat)
The Nobel Prize in Chemistry 1907
"for his biochemical
researches and his discovery
of cell-free fermentation"

Eduard Buchner
Germany
Landwirtschaftliche Hochschule
(Agricultural College) Berlin,
Germany
b. 1860, d. 1917.
5
Enzim mempercepat Laju
Reaksi
 Bagaimana agar molekul bergerak lebih cepat?

•Sistem biologis sangat sensitif terhadap

perubahan suhu, pH, dan lingkungan

•Enzim dapat mempercepat laju reaksi tanpa

mengalami perubahan suhu

•Dengan cara menurunkan Energi Aktifasi

Reaksi

•Melalui jalur reaksi yang baru “short cut”

 Energi Aktifasi
adalah energi
yang diperlukan
untuk memulai
jalannya reaksi.
 Energetika Reaksi Enzimatik

 Reaksi dengan Enzim berlangsung 108 - 1011 kali lebih cepat

dibandingkan reaksi tanpa enzim
Mekanisme Reaksi Enzim

enzim
A+B P+Q
Substrat Produk
 SPESIFISITAS
• Enzim sangat spesifik. Tiap enzim hanya
mengkatalis satu tipe reaksi dan terkadang hanya
bekerja dengan satu substrat spesifik

◦ Ex. NH2-C-NH2 + H20 urease 2NH3 + CO2
O

• Urease tidak memiliki efek apapun pada seny. lain

 Kelas Enzim
 Spesifisitas Absolut
 hanya bereaksi dengan satu jenis substrat

 Spesifisitas Gugus
 Bekerja dengan sejumlah molekul dengan
gugus fungsi yang sama

 Spesifisitas Stereokimia
 Hanya bekerja pada struktur D- atau L-
Sisi Aktif Enzim dan Substrat
• Gaya Van der Waals
• Gaya elektrostatik
(interaksi ionik)
• Ikatan Hidrogen
• Interaksi Hidrofobik

enzim
A+B P+Q
Substrat Produk
 Kompleks Enzim-substrat
 tahap 1:
E + S ES
 Enzyme Substrate Complex

+
 Kompleks Enzim-substrat
 Tahap 2:
.

 ES EP

ES transition EP
state
 Kompleks Enzim-produk
 Tahap 3 :

 EP E + P
The product
is made
Enzyme is
ready
EP for
another
substrate.
 Mekanisme Reaksi Enzimatis
 “Lock and Key” : Emil Fischer (1894)
 lock = enzim, key = substrat

a-metilglukosida
diketahui dihidrolisis oleh
invertin, tetapi tidak oleh
emulsin,

b-metilglukosida
dihidrolisis oleh emulsin,
tetapi tidak oleh invertin:

Enzim dan glukosida

komplemen satu sama
lain seperti sebuah
kunci dan gemboknya
(kompleks Enzim-Substrat)
 Mekanisme Reaksi Enzimatis
 “Induced Fit” : Daniel Koshland (1958)

Heksokinase :
(a) Tanpa (b) dengan substrat glukosa
http://www.biochem.arizona.edu/classes/bioc462/4
Enzim

Pemotongan
(degradasi)

Penyusunan
(sintesis)

Perubahan
(modifikasi)
 Klasifikasi Enzim
Berdasarkan tipe reaksi yang dikatalisisnya :
Oksidoreduktase : mengkatalis reaksi redoks
Transferase : pemindahan gugus fungsi

Hidrolase : mengkatalis reaksi hidrolisis

Liase : +/- ikatan C=C

Isomerase : penataan membentuk isomer

Ligase : penggabungan dua molekul

 Sisi Aktif Enzim

 Suatu lokasi pada enzim:

sisi aktif, yang
berinteraksi dengan
substrat

© H.PELLETIER, M.R.SAWAYA
ProNuC Database
 Substrat
 Substrat dari suatu enzim adalah
Reaktan yang diaktifkan oleh enzim
 Enzim spesifik terhadap substratnya
 Ditentukan oleh sisi aktif enzim

Substrat : Trigliserida  Gliserol + 3 Asam Lemak

E + S E S E P E + P

© 2007 Paul Billiet ODWS

Rate enhancement
Fe3+ →1000 fold
Hemoglobin → 1 ,000,000 fold
Catalase → 1 ,000,000,000 fold
2 H2O2 → 2 H2O + O2
200,000 catalytic events/second/subunit
(near the diffusion-controlled limit).
The reaction is sped up by a billion fold!

(a prosthetic group)

Active site
(tetramers)
Each enzyme has at least one active site
 Kofaktor dari suatu Enzim
 Bagian non-protein dari
suatu enzim

 Kofaktor :
 1. Anorganik : Mg2+, Mn2+
 2. Organik (Koenzim)

 Koenzim yang terikat

kuat secara kovalen :
gugus prostetik

 banyak vitamin
merupakan koenzim

Nitrogenase enzyme with Fe, Mo and ADP cofactors

Jmol from a RCSB PDB file © 2007 Steve Cook
Apoenzim + Kofaktor = Holoenzim
H.SCHINDELIN, C.KISKER, J.L.SCHLESSMAN, J.B.HOWARD, D.C.REES
STRUCTURE OF ADP X ALF4(-)-STABILIZED NITROGENASE COMPLEX AND ITS

IMPLICATIONS FOR SIGNAL TRANSDUCTION; NATURE 387:370 (1997)

Contoh Sejumlah Kofaktor :
 Vitamin sebagai bahan baku Koenzim

Vitamin Koenzim Deficiency Disease

Biotin Biocytin Not observed

Cobalamin (B12) Cobalamin Pernicious anemia
Folic acid tetrahydrofolate Megaloblastic anemia

Nicotinamide Nicotinamide Pellagra

Pantothenate Coenzyme A Not observed
Pyridoxine (B6) Pyridoxal phosphate Not observed
Riboflavin (B2) Flavin Not observed
Thiamine (B1) Thiamine pyrophosphate Beriberi
The Nobel Prize in Chemistry 1946

Sumner, J. B. (1926) “ The

isolation and crystallization
of the enzyme urease” J.
Biol. Chem. 69:435-441.

Cornell University
Ithaca, NY, USA
 Koenzim – senyawa organik non protein organik
 Koenzim – senyawa organik non protein organik
Karbonik Anhidrase

- 
CO 2  H 2O  HCO3  H
Pengaruh Suhu thdp. Rx. Enzimatik

Suhu Optimum
umumnya
Laju Reaksi

37oC.

Suhu
 Efek pH terhadap Rx. Enzimatik

Umumnya enzim
Bekerja Optimum
pada pH 7.4
Reaction Rate

Tidak semuanya

pH
Contoh pH Optimum Enzim
 Enzim sumber pH Optimum

 pepsin gastric mucosa 1.5

 sukrase intestine 6.2
 katalase liver 7.3
 arginase beef liver 9.0
 alkalin bone 9.5
 PROENZYMES (ZYMOGENS)
 Enzymes manufactured in inactive form.
Activated when small part
of polypeptide chain removed.

Hormones,
Digestive Enz,
Blood Clotting Enz
 PROENZYMES (ZYMOGENS)
Enzymes manufactured in inactive form.
In pancreas (inactive) In blood (active)
Proinsulin Insulin

S S
S S S S
S S
S S
S S
 PROENZYMES (ZYMOGENS)
(inactive) (active)
In pancreas In Intestines
enteropeptidase Trypsin
Trypsinogen

Chymotrypsinogen Trypsin Chymotrypsin

procarboxypeptidase Trypsin Carboxypeptidase

Digestive Proteases
Enzymes Cleave peptides
 Some Enzyme Terminology
• Enzyme –
•
• Substrate –

• Enzyme Specificity –

• Cofactor –

• Coenzyme –
 Some Enzyme Terminology
• Enzyme – a biomolecule that catalyzes biochemical
reaction by lowering activation energy

• Substrate – the substance that undergoes a chemical

change by an enzyme

• Enzyme Specificity – the characteristic that an enzyme

acts on related substrates

• Cofactor – a nonprotein molecule or ion required by an

enzyme for catalytic activity

• Coenzyme – an organic molecule required by an

enzyme for catalytic activity
More Enzyme Terminology
• Apoenzyme – a catalytically inactive protein formed by
removal of the cofactor from an active enzyme

• Holoenzyme – apoenzyme + cofactor

• Active Site – the location on an enzyme where a substrate is

bound and catalysis occurs

• Enzyme Activity – the rate at which an enzyme catalyzes a

reaction

• Optimum Temperature – the temperature at which enzyme

activity is highest
• Optimum pH - the pH at which enzyme activity is highest
• Zymogen (proenzyme) – the inactive enzyme precursor
Enzyme Inhibitors
 Competive - mimic substrate, may block active site, but may dislodge it.
 Enzyme Inhibitor
 They are molecules that bind to
enzyme ,either at its active site or
else where, and cause reduction in
enzyme activity.
 The inhibitor may bind to enzyme
reversibly or irreversibly

44
 Coenzyme NAD+

 NAD+ transports electrons from one

metabolic pathway to another
 Drugs as Enzyme Inhibitors
 Drug mimics some natural chemical to block enzyme.
 Drugs as Enzyme Inhibitors
 Drug mimics some natural chemical to block enzyme.
Applications of Proteolytic Enzymes
 Dairy:
 Calf rennet (chymosin) is used in the coagulation of milk protein for
cheese production, without loss of sensitive components.
 Lactase hydrolyzes the principal carbohydrate of milk, lactose. This
processes a cheese byproduct and relieves lactose intolerance.
 Detergents:
 Protein stain removal is facilitated by the hydrolysis of proteins into
oligopeptides. Enzyme stability with respect to storage, pH, temperature
and bleach are key concerns.
 Leather Production:
 Proteases are widely used for the soaking and dewooling stages of hide
processing in which selective protein degradation results in a softer
produce without substantial loss of strength.

 Food and Feed:

 Starch conversion to high-fructose corn syrup is an important process to
the beverage industry.
17.4
CHEE 323 8
 Fungal Enzymes
Enzyme EC Sources Application

a-Amylase 3.2.1.1 Aspergillus E Baking

Catalase 1.11.1.6 Aspergillus I Food
Cellulase 3.2.1.4 Trichoderma E Waste
Dextranase 3.2.1.11 Penicillium E Food
Glucose oxidase 1.1.3.4 Aspergillus I Food
Lactase 3.2.1.23 Aspergillus E Dairy
Lipase 3.1.1.3 Rhizopus E Food
Mucor
Rennet 3.4.23.6 E Cheese
miehei
Pectinase 3.2.1.15 Aspergillus E Drinks
Protease 3.4.23.6 Aspergillus E Baking

E: extracellular enzyme; I: intracellular enzyme

 Bacterial Enzymes

Enzyme Sources Application

a-Amylase 3.2.1.1 Bacillus E Starch

b-Amylase 3.2.1.2 Bacillus E Starch
Escherichia
Asparaginase 3.5.1.1 I Health
coli
Glucose Fructose
5.3.1.5 Bacillus I
isomerase syrup
Penicillin Pharmace
3.5.1.11 Bacillus I
amidase utical
Protease 3.4.21.14 Bacillus E Detergent
Penicillin: War of Enzyme against
Enzyme.
• Produced by mold, it prevents growth of bacteria by
successfully competing for active sites on an enzyme
that bacteria need for cell wall production.
 1. Bacteria need the enzyme transpeptidase
to make their cell walls rigid and cross-linked.
 2. Penicillin takes control of transpeptidase.
 3. Bacteria cell walls are not cross-linked and
the contents of the bacteria cells cannot be
held in.
 4. Cytoplasm spills out, and the bacteria die.
By changing the R group, science has found a
way to prevent this from happening.