BIOKIMIA ENZIM

ENZYMES
Pendahuluan (overview) Enzyme Specificity Kofaktor Enzim Enzyme Nomenclature Lock and Key Enzyme-Substrate Complex Induced Fit Theory Cara Kerja Enzim Conditions Affecting the Actions of Enzymes Inhibitor Enzim Isozim/Isoenzim Regulation of Enzymatic Activity

PENDAHULUAN
Organisme hidup rangkaian reaksi biokimia peran enzim Enzim : biokatalisator yg mengatur kecepatan berlangsungnya semua proses fisiologis Hampir semua dr 2000/ lebih enzim yg diketahui merupakan protein globular Aktivitas katalitiknya bergantung pd integritas struktur sbg protein

Contoh : Jika enzim dididihkan dg asam kuat/diinkubasi dg tripsin (perlakuan yg memotong rantai polipeptida) aktivitas katalitiknya akan hancur pentingnya struktur kerangka primer protein enzim

Jika struktur berlipatnya rantai protein yang khas dr enzim diubah (oleh panas, pH extrim, senyawa perusak ) aktivitas katalitiknya juga lenyap jadi struktur primer, sekunder dan tertier protein juga penting bagiaktivitas katalitiknya.

KESPESIFIKAN ENZIM

Enzymes are highly specific both in the reactions they catalyze and in the compounds (reactants/substrates) on which they act Contoh : enzim proteolitik Mengkatalisis hidrolosis ikatan peptida Enzim2 proteolitik berbeda tingkat spesifisitas substratnya, misalnya :
SUBTILIN pd bakteri tdk bedakan ik. peptida yg akan diputus TRIPSIN ik. pep pd sisi karboksil arginin/lisin TROMBIN ik.pep antara arginin dan glisin

KOFAKTOR ENZIM
Dlm fungsinya sbg katalisator suatu reaksi kadang enzim cukup mengandalkan struktur proteinnya, tp. kadang enzim butuh senyawa lain yang bukan protein yang disebut KOFAKTOR Yg termsk kofaktor :

ION LOGAM MAUPUN SUATU MOLEKUL ORGANIK KOMPLEKS YG DISBT KOENZIM KOENZIM/GUGUS YG MEMBENTUK IKATAN KOVALEN DG ENZIM DISBT SBG GUGUS PROSTETIK.

Kompleks enzim-kofaktor disebut Holoenzim, Jikakofaktornya diambil, disebut Apoenzim

ENZYMES

Enzymes are PROTEIN CATALYSTS

Large complex proteins made up of one or more polypeptide chains In Vivo called an enzyme. Outside the body it is just called a catalyst Control the rate of chemical reactions that take place in cells, tissues, and organs.

Each chemical reaction that occurs in a living system requires the assistance of a specific enzyme

Catalyst: A substance that changes the speed of a chemical reaction without itself undergoing a permanent chemical change in the process

OVERVIEW

In a chemical reaction, reactants collide and enter into a transition state

this state is short and rapidly breaks down to either products or reactants

Free energy (G) Free Energy of Activation ( G ) Free Energy difference ( G)

WHAT IS HAPPENING...
Catalyst

reduces amount of activation energy so the reaction occurs faster Catalyst accelerates both the forward and reverse reactions and thus only increases the rate at which a reaction approaches equilibrium

Endoenzyme Found and used within the cell Exoenzyme Secreted into extracellular environment

later taken up by the cells for use

ENZYME NOMENCLATURE

The official name of an enzyme has TWO (2) parts

First: names the substrates or the products of the reaction Second: Designates the type of reaction catalyzed

CLASSES OF ENZYMES

NAMING AND CLASSIFICATION

Enzymes can be classified by chemical reaction catalyzed 1. Addition/Removal of water a) Hydrolase - carbohydrase, amidase,

protease
b) Hydrase -fumarase, enolase, carbonic

anhydrase

2.

Transfer of electrons

a)
b)

Oxidase
Dehydrogenase

3.

Transfer of a radical a) Transglycosidase - monosaccharides Transphosphrylase & Phosphomutase - phosphate group Transaminase - amino group Transmethylase - methyl group Transacetylase - acetyl group

b)
c) d) e)

4.

Splitting of forming a C-C bond a) Desmolase Changing geometry or structure of molecule a) Isomerase
Joining two molecules through hydrolysis of pyrophosphate bond in ATP of tri-phosphate a) Ligase

5.

6.

LOCK AND KEY THEORY

Lock and Key theory: simple analogy commonly used to explain the specificity of enzymes A specific key will only open a specific lock The key can be used over and over on the same type of lock

ENZYME-SUBSTRATE COMPLEX
Explains the specificity of enzyme action Substrates of an enzymatic reaction bind to a specific site on the enzyme shape of that site is complementary to that of the substrates Active Site: part of an enzyme to which the substrates bind (most cases: a pocket of groove in the surface of the protein) Included within the pocket are functional groups that attract the substrate molecules and mediate the catalytic event

ENZYME-SUBSTRATE COMPLEX DRAWING

EXAMPLES OF ACTIVE SITES

INDUCED FIT THEORY

Active site on enzyme not as rigid as lock and key model As the substrate attaches to the enzymes active site, the site changes shape to fit the substrate improves the fit of the active site to the substrate brings catalytic groups into the correct position for action

MECHANISMS OF CATALYSIS
A

large part of the catalytic power of an enzyme depends on its ability to lower the activation energy To do so, an enzyme may provide an environment within the active site that favors the transitions state...or it may provide catalytic groups that allow the reaction to proceed via intermediates not part of the uncatalyzed reaction many enzymes act as general acid-base catalysts

MECHANISMS OF CATALYSIS

Polypeptides cannot by themselves catalyze all of the biologically important reactions.

Use coenzymes and cofactors

COFACTORS / COENZYMES
Cofactors:

non-protein component of enzymes required for the enzyme to function inorganic like a cofactor, but organic found mostly in vitamins

Coenzymes:

Both

cofactors and coenzymes make up part of the active site

COENZYMES

Interacts with the active site of the enzyme and acts as a catalytic group in the reaction

some are permanently attached

either covalently or non covalently

Each type of coenzyme is specialized to perform one of a small number of biochemical functions but may perform that same function for more than one enzyme

BIOTIN: EXAMPLE OF COENZYME

Serves as the coenzyme for several enzymes one of the 9 watersoluble vitamins required by humans Example: pyruviate carboxylase acetyl-CoA carboxylase

both catalyze the fixation of Co2

CONDITIONS AFFECTING THE ACTIONS OF ENZYMES

Heavy Metal Ions Temperature pH salt Amount of Substrate concentration

*everything

that affect a protein affect an enzyme because ENZYMES ARE PROTIENS

HEAVY METAL IONS

Can disrupt enzymatic activity

when present at active site, substitution of a different metal ion for the original ion causes the enzyme to malfunction and denature

EXTREMES OF TEMPERATURE

Changes enzyme structure changes active site

prevents enzyme from attaching to substrate

The thermal agitation of the enzyme molecule disrupts the hydrogen bonds, ionic bonds, and other weak interactions within the protein molecule In humans, enzymes have an optimum temperature of 37C

PH

CHANGES ON ENZYME ACTIVITY

Extreme changes in pH values denature such ionisable enzymes rendering them ineffective

within a narrow pH range, enzyme structure changes reversibility, and each such enzyme was optimally at a specific pH

Optimal pH values for most enzymes

6-8 pH exception: I.e. digestive enzymes in stomachs

INHIBITORS
Competitive

Inhibitors

Compete with substrate for the enzymes Do not affect Vmax Raise the apparent Km

NONCOMPETITIVE INHIBITORS
Do

not affect Km Lower Vmax Slows down dissociation of ES

ENZYMES IN BIOTECHNOLOGY
Biotechnology

is the application and harnessing of microorganisms (such as bacteria, viruses and fungi) or biological processes to produce desired and useful substances and facilitate industrial processes.

FERMENTATION: EXAMPLE OF BIOTECHNOLOGY

Ex:

The manufacture of wine, involves the fermentation of grape juice, a rich source of glucose, by wild yeasts present on grape skin

fermentation of sugar by yeast is the basis for the production of other alcoholic drinks

Better

brewing through improving yeast and large scale production are the results of biotechnology

genes responsible for the yeast enzymes have been cloned and bacteria has been used in large scale production of the yeast enzymes needed

GENETIC ENGINEERING
Involves the manipulation of genes used to describe the modern techniques in molecular biology where genes can be removed from one type of a cell to another, altering a cells properties USED IN: producing new antibodies, insulin, and biological detergents

BIOLOGICAL DETERGENT
Lipolase:

enzyme which is constituent of biological detergents, consists of fatdigesting (splitting) enzymes Widely used in the soap and detergent industry

save energy rapidly biodegradable, thus leaving no harmful residues no negative impact on sewage treatment processes pose no risk to aquatic life

STREPTOKINASE IN BREAKING DOWN BLOOD CLOTS

STREPTOKINASE

can dissolve blood clots that form in the heart, blood vessels, or lungs after a process, such as a heart attack called a thrombolytic agent can also dissolve blood clots that form in intravenous catheters

tubing that goes into a vein for fluid exchange