ENZYME- BIOLOGICAL CATALYST - PowerPoint PPT Presentation

1 / 48
About This Presentation
Title:

ENZYME- BIOLOGICAL CATALYST

Description:

BIOLOGICAL CATALYST Enzyme As Catalyst All enzymes are proteins - with the exception of some RNAs that catalyze their own splicing all enzymes are proteins In general ... – PowerPoint PPT presentation

Number of Views:762
Avg rating:3.0/5.0
Slides: 49
Provided by: BillB216
Category:

less

Transcript and Presenter's Notes

Title: ENZYME- BIOLOGICAL CATALYST


1
ENZYME-BIOLOGICAL CATALYST
2
Enzyme As Catalyst
  • All enzymes are proteins - with the exception of
    some RNAs that catalyze their own splicing all
    enzymes are proteins
  • In general, names end with suffix ase
  • - tyrosinase (tyrosine), celullase (cellulose),
    protease (protein), lipase (lipid)
  • Enzyme a biological catalyst
  • enzymes can increase the rate of a reaction by a
    factor of up to 1020 over an uncatalyzed reaction

3
  • Catalysis
  • - the process of increasing the rate of chemical
    reactions
  • Catalyst
  • - the substance that facilitate in catalysis

4
Enzyme As Catalyst
  • Enzymes are catalysts
  • - increase the rate of a reaction
  • - not consumed by the reaction
  • - act repeatedly to increase the rate of
    reaction
  • - enzymes often specific promote only 1
    particular reaction, others catalyze a family of
    similar reactions
  • cellulase cellulose as substrate
  • hexokinase any 6 ring monosaccharide -
    fructose, glucose

5
General Properties of Enzyme
  • Higher reaction rates
  • - the rates of enzymatically catalyzed reactions
    are 106 to 1020 gt than uncatalyzed reaction
  • Milder reaction rates
  • - enzyme catalyzed reactions occur under
    relatively milder conditions lt 100oC,
    atmospheric pressure and nearly neutral pH-
    contrast with chemical catalysis requires high
    temperature, pressures and extremes pH.
  • Greater reaction specificity
  • - enzyme have greater degree of specificity to
    their substrates and their products rarely have
    side products

6
Enzyme Catalysis
Active site - part of enzyme to which the
substrate binds and the reaction takes place
Substrate a reactant in an enzyme-catalyzed
reaction
Product
Enzyme-substrate (ES) complex the intermediate
formed when the substrate is bind at the active
site of an enzyme
7
Enzyme Catalysis
  • GENERAL FORMULA

E enzyme S substrate P product
8
Enzyme catalysis reaction
  • Physically interact with their substrates to
    effect catalysis
  • E S ES ES EP E P
  • Where
  • - E enzyme
  • - ES enzyme/substrate complex
  • - ES enzyme/transition state complex
  • - EP enzyme/product complex
  • - P product

9
Enzyme catalysis reaction
  • Substrate bind to the enzymes active site
  • pocket in the enzyme
  • Catalytic site active site where reaction
    takes place

10
Enzyme catalysis reaction
  • E S ES ES EP E P
  • 1st step enzyme binds to substrate molecule to
    form an enzyme substrate complex
  • E S ES

Enzyme
11
Enzyme catalysis reaction
  • E S ES ES EP E P
  • 2nd step Formation of the transition state
    complex where the bound substance is neither
    product nor reactant
  • ES ES, ES?ES

12
Enzyme catalysis reaction
  • E S ES ES EP E P
  • 3rd step Formation of the enzyme product
    complex ES EP

13
Enzyme catalysis reaction
  • E S ES ES EP E P
  • 4th step Release of product EP E P

14
Enzyme catalysis reaction
  • Enzyme can only work on one substrate molecule at
  • a time
  • Not change during the reaction
  • One product is release, enzyme is available to
    accept another substrate molecule

15
Enzyme Catalysis
  • Rate of reaction reaction velocity (V)
  • - the rate of enzyme reaction is measured by the
    rate of the appearance of products or the rate of
    disappearance of substrates.
  • - dP/dT or dS/dT
  • ?mol product/min or ?mol substrate/min
  • Enzyme activity?
  • 1 unit (U) is the amount of enzyme that catalyses
    the reaction of 1 ?mol of substrate per minute
    under specified conditions.

16
Enzyme Catalysis
  • The rate of a reaction depends on its activation
    energy, DG
  • an enzyme provides an alternative pathway with a
    lower activation energy
  • Activation energy the energy required to start
    a reaction
  • Transition state the intermediate stage in a
    reaction in which the old bonds break and new
    bonds are formed

17
How enzyme work?
  • Transition state theory
  • The enzyme (E) must approach the substrate (S),
    the substrate attach to the active site through
    noncovalent bond
  • Formed the high energy (unstable) ES complex
  • In ES complex, the covalent bond in substrate is
    in the process of breaking while the EP complex
    is forming.

18
Enzyme Catalysis - Example
  • Consider the reaction

catalase
  1. No catalyst,
  2. with added Fe3 salt,
  3. with added catalase

19
  • (a) a/e for the reaction in the absence of a
    catalyst
  • (b) a/e lowered in the presence of an iron
    catalyst
  • (c) energy diagram for the catalase-catalyzed
    breakdown of H2O2
  • (d) energy diagram for the noncatalysed
    breakdown of H2O2 at elevated temperature

a/e activation energy
20
Active site
  • Has specificity can discriminate among possible
    substrate molecules
  • - others recognize a functional group (group
    specificity)
  • - only recognize one type of molecule (eg. D vs
    L isomer)
  • (absolute specificity)
  • Relatively small 3D region within the enzyme
  • - small cleft or crevice on a large protein
  • Substrates bind in active site by weak
    non-covalent interactions (Hydrogen bond,
    hydrophobic and ionic interaction)

21
Active site
  • The interactions hold the substrate in the proper
    orientation for most effective catalysis
  • The energy derived from these interactions
    binding energy
  • Binding energy is used, in large part to lower
    the activation energy and stabilize the
    transition state complex (ES)
  • Each non-covalent interaction provides energy to
    stabilize the transition state

22
Binding Models
  • Two models have been developed to describe
    formation of the enzyme-substrate complex
  • Lock-and-key model substrate binds to that
    portion of the enzyme with a complementary shape
  • Induced fit model binding of the substrate
    induces a change in the conformation of the
    enzyme that results in a complementary fit

23
Enzyme/substrate interaction
  • Lock and key model
  • - substrate (key) fits into a perfectly shaped
    space in the enzyme (lock)
  • - lots of similarities between the shape of the
    enzyme and the shape of the substrate
  • - highly stereospecific
  • - implies a very RIGID inflexible active site
  • - site is preformed and RIGID

24
Enzyme/substrate interaction
  • Induced fit model (hand in glove analogy)
  • - count the flexibility of proteins
  • - substrate fits into a general shape in the
    enzyme, causing the enzyme to change shape
    (conformation) close but not perfect fit of E
    S
  • - change in protein configuration leads to a
    near perfect fit of substrate with enzyme

Figure 6.3, pg 148 Campbell Farrell,
Biochemistry, 6th Ed., 2009, Thomson Brooks/Cole
25
Key words for today
  • You need to understand
  • Factors effecting enzyme reaction rate
  • 6 classes of enzymes
  • Cofactor, coenzymes, holoenzymes, apoenzymes
  • Allosteric enzyme, effectors (positive and
    negative), heterotropic and homotropic
    allosterism
  • Isoenzyme and multienzyme

26
Characteristics of enzyme reactions
  • What influence the enzyme reaction rate?
  • Substrate concentration
  • Temperature
  • pH
  • Enzyme concentration
  • Inhibitor

27
Characteristics of enzyme reactions
  • Substrate Saturation Increasing the substrate
    increases the rate of reaction (enzyme activity).
  • enzyme saturation limits reaction rates. An
    enzyme is saturated when the active sites of all
    the molecules are occupied most of the time.
  • At the saturation point,
  • the reaction will not speed
  • up, no matter how much
  • additional substrate
  • is added. The graph of
  • the reaction rate will plateau.

substrate substrate concentration
28
Characteristics of enzyme reactions
  • Temperature
  • - very sensitive to temperature changes
  • - low temp, rate of an enzyme-catalysed reaction
    increases proportionally with increasing
    temperature

29
Characteristics of enzyme reactions
  • Effects of Temperature
  • All enzymes work within a range of temperature
    specific to the organism.
  • Increases in temperature lead to increases in
    reaction rates - is a limit to the increase
    because higher temperatures lead to a sharp
    decrease in reaction rates - due to the
    denaturating (alteration) of protein structure
    resulting from the breakdown of the weak ionic
    and hydrogen bonding that stabilize the three
    dimensional structure of the enzyme.

30
Characteristics of enzyme reactions
  • pH
  • - enzymes have an optimal pH at which they
    function properly
  • - varies to each other but most in the range of
    pH 6-8

pepsin in the stomach works best at a pH of 2 and
trypsin at a pH of 8.
31
Characteristics of enzyme reactions
  • Effects of pH Most enzymes are sensitive to pH
    and have specific ranges of activity.
  • All have an optimum pH. The pH can stop enzyme
    activity by denaturating (altering) the three
    dimensional shape of the enzyme by breaking
    ionic, and hydrogen bonds.

32
Characteristics of enzyme reactions
  • Enzyme concentration
  • - the higher the concentration, the greater
    should be the initial reaction rate will be
    lasting as long as substrate present

33
Characteristics of enzyme reactions
  • Inhibitor
  • - inhibit enzyme by occupy the active site or
    bind to other part of enzyme leading to the
    change of enzyme shape and eventually the active
    site
  • - this will decrease the enzymatic reaction rate

34
Classification of Enzymes
  • Have 6 categories
  • Each enzyme has an official international name
    ending with ase and a classification number
  • Number consists in 4 digits (referred to a class
    and subclass of reaction

35
Classification of Enzymes
36
Classification of Enzymes
Table 5.1, pg 136 Boyer, R., Concepts in
Biochemistry, 3rd Ed., 2006, John Wiley Sons
37
Enzyme Classes Examples
Class Example Reaction
1 (oxidoreductase) alcohol dehydrogenase
2 (transferase) hexokinase glucose ATP ? glucose-6-phosphate ADP
3 (hydrolase) chymotrypsin polypeptide H2O ? peptides
More examplesRefer Table 5.2, pg 136 , Boyer,
R., Concepts in Biochemistry, 3rd Ed., 2006, John
Wiley Sons
38
Enzyme Classes Examples
Class Example Reaction
4 (lyase) pyruvate decarboxylase
5 (isomerases) alanine racemase D-alanine L-alanine
6 (ligases) pyruvate carboxylase
39
Enzymes cofactor
  • Enzymes require chemical entity in order to
    function properly (assists an enzyme in catalytic
    action)
  • Cofactor nonprotein molecule that assist in an
    enzyme catalytic reaction
  • Coenzyme smaller organic or organometallic
    molecule derived from vitamin, weakly bound to
    enzyme, temporarily associated with enzymes
  • Prosthetic group coenzymes that are covalently
    or noncovalently tightly bound to enzyme and
    always present.

40
Enzymes cofactor
  • Holoenzyme an enzyme in its complete form
    including polypeptide(s) and cofactor
  • Apoenzyme enzyme in its polypeptide form
    without any necessary prosthetic groups or
    cofactors

41
Allosteric Enzymes
  • Allosteric enzyme an oligomer whose biological
    activity is affected by other substances binding
    to it
  • these substances change the enzymes activity by
    altering the conformation(s) of its 4structure
  • Allosteric effector a substance that modifies
    the behavior of an allosteric enzyme may be an
  • allosteric inhibitor negative effectors
  • allosteric activator positive effectors

42
Allosteric enzyme in feedback Inhibition
  • Formation of product inhibits its continued
    production feedback inhibition

43
Allosteric Enzymes (Contd)
  • The key to allosteric behavior is the existence
    of multiple forms for the 4structure of the
    enzyme
  • allosteric effector a substance that modifies
    the 4 structure of an allosteric enzyme
  • homotropic effects allosteric interactions that
    occur when several identical molecules are bound
    to the protein e.g., the binding of aspartate to
    ATCase
  • heterotropic effects allosteric interactions
    that occur when different substances are bound to
    the protein e.g., inhibition of ATCase by CTP
    and activation by ATP

ATCase aspartate transcarbomylase CTP
cytidine triphosphate
44
Allosteric enzymes
  • A change in conformational structure at one
    location of a multisubunit protein that causes a
    conformational change at another location on the
    protein
  • Effectors i) serves as stimulants to enzyme
    (ve effectors) increase catalytic
    activity ii) inhibitors (-ve effectors) to
    enzyme
    reduce/inhibit catalytic activity
  • - Act by reversible, noncovalent binding to a
    site on the enzyme
  • Larger and more complex than nonallosteric enzyme
  • Have 2 or more subunits (oligomeric)
  • Allosteric enzymes have regulatory sites for
    binding of substrates and reaction (catalytic
    sites)

45
HOMOTROPIC ALLOSTERISM
  • Eg. Tetrameric allosteric enzyme composed of 4
    identical subunits
  • Each subunit has a catalytic site where
    substrate/effector will bound and transformed to
    product
  • Once bound to active site, a message will
    transmitted via conformational changes to an
    active site on another subunit which makes it
    easier for a substrate molecule to bind and react
    at that site
  • This type (substrate and effector) are the same
    is called cooperative or homotropic

46
HETEROTROPIC ALLOSTERISM
  • A dimer with nonidentical subunits
  • Subunit a contain the active site catalytic
    subunit
  • Subunit ß contains the site for effector binding
    regulatory subunit
  • Binding of a specific effector molecule to the
    regulatory site on the ß subunit sends a signal
    via conformational changes to the catalytic site
    on subunit a
  • Substrate and effector different kinds of
    molecules - heterotropic

47
Isoenzymes
  • Enzymes that catalyze the same reaction
    (catalytically and structurally similar) but are
    encoded by different genes
  • Glycogen phosphorylase-synthesize in liver, brain
    and muscle-involves in degradation of glycogen
  • Isoenzymes isoforms

48
Multienzymes
  • A group of noncovalently associated enzymes that
    catalyze 2 or more sequential steps in
    metabolic/biochemical pathway
Write a Comment
User Comments (0)
About PowerShow.com