1 Enzyme Intro

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1.ENZYME ENGINEERING


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= IN YEAST1878 Khne

Special groups of proteins and their biological functions:

RegulRegul aatortor proteins proteinslac-repressor RNA synthesisinterferons vrus resistencyinsuline glucose metabolism

growth hormoneTransport proteins

lactose permease cell membrane transportmyoglobin O 2 - in muscle

hemoglogin O 2 in bloodProtecting proteins

antibodies (abzymek) foreign material -komplexthrombine blood clotting


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Toxins B.thuringiensis biol. InsekticideCl.botulinum food poisoning

Reserve proteinsovalbumin egg-whitekasein milk proteinzein corn (maize) germ

Contractile proteinsdynein cilia, flagella

Structural proteinscollagen joints, tendonsglycoproteins cell wall

ENZYMES catalists of reactions

chaperonok prionok


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Catalitic effect

RegulRegul aatortor proteins proteins

lac-repressor RNS synthesisinterferons vrus resistencyinzuline glkse metabolismgrowth hormone

Transport proteinslaktose permease cell membrane transportmyoglobine O 2 - in muscle

hemoglogine O 2 in blood

Protecting proteinsantibodies (abzymek) foreign material -komplex

thrombine blood clotting


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A + B AB* P

*ST.*GH* +=

BA

ABCC

CK =

hkT

CCCk dtdP

ABBAr ==

kThk

CCC

K r

BA

AB

==

**r ** STHkT

hk RTlnRTlnK G ===

RTE

RTH

R S

r ekonsteekTk

=

T az abszolt hmrsklet (Kelvin fok)

k a Boltzmann lland (1,37.10 -16 erg/ oC)h a Planck lland ( 6,62.10 -27)

( ) ( )katnemkat EE

( ) ( )katr nemkatr k k

b

Eyring: in 1930s, transition state theory


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ACTIVATED COMPLEX ES*

FUNCTIONAL DOMAINS

SUBSTRATE BINDING SITE

REGULATOR DOMAINS:Memodulator (INHIBITOR,ACTIVATOR,S,P)

Covalent modification:

phosphorilationglycosylation proteolysis

Catalytic domain ACTIVE CENTER

They could beidentical, too


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SUBSTRATE BINDING SITESack, pocket on the surfaceElectrostatic and Hydrofobic interactions

noncovalent interactions

Only a small portion of the surface of the prot molecule

General cases of the enzymatic catalysis: 1. acid-base

modern theory Linus Pauling 1946 2.kovalent catalysisHaldane 1930 3.metal ion catalyis

Effects, responsible for and characterize the enzyme-catalysis: proximity effect

orientation effect lock and key modelinduced fit Daniel Koshland-conformation change

ENTHROPY TRAPREACTIV GROUPS

PROTEINSTRUCTURE

22


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ENERGIAGTENERGY BARRIER


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S

szabad E

+

ES komplex

szabad E

+termkek

KULCS

ZR

-

LOCK AND KEY MODELL

r ent c s e e tus


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r ent c s e e tus

A sztereospecifits alapj

ORIENTATION EFFECT

Basis of stereospecificity


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http://www.chem.ucsb.edu/~molvisual/ABLE/induced_fit/index.html


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, ,

ESS

S

G*nemkat

G*kat

P

ES

P

A COO REACTIVE GROUPS


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Asp -COO-Cys -SHGlu -COO- & -CONH2

His HC C imidazole

+HN NHCH

Lys - NH2

Met CH3 S -Ser -OH

Thr CH3CHOH -

terminal -NH2 s -COO-

electrostatic klcsnhats

H - bondsC O.....H O

C O.....H N

REACTIVE GROUPS...


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Structure of proteins


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Formation of the active center


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ADVANTAGES OF ENZYMES

HIGHER REACTION VELOCITIES 106

-1012

*MILDER REACTION CIRCUMSTANCESHIGHER REACTION -..+ SPECIFICITYREGULATION IS POSSIBLE

PROPERTIES OF ENZYMES AND E-CAT. REACTIONSTHERM.DYNAMICALLY POSSIBLE RECTIONS

G


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1~10 2~ 10 7

150 27030 5016,7

-Cu2+

lipoxigenase

Linoleic acid + O 2 ?linoleicperoxid

15,6 x 10 10

10746

H+

invertaseSacharose + H 2O ?

glucose+fructose

1

2,1 x 10 686

50

H+

tripsin

Cazein + nH 2O

? (n+1)peptid

12,1 x 10 3

3,5 x 10 8

7556,526,8

-I-

catalase

H2O2? H2O + 1/2O 2

k relative25 oC

ActivationenergykJ/mol

Catalysator

Reaction


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APOENZYME + COENZYME prostetic group (covalent)COFACTOR

HOLOENZYME

PROTEIN


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HOLOENZYME


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APOENZYME + COFACTOR

HOLOENZYME

(IN)ACTIVE PROTEIN

METALMg,Ca,Zn,Fe,Cu,Mo

KOENZYME

Prostetic groupstable covalent bound.

FAD(H 2), Hem,Piridoxal-P(B 6)

CosubstrateCyclic regeneration nee

NAD(H), ATP

NOMENCLATURE OF ENZYMES


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1. Substrate urea + water CO 2 + 2NH 3

urease

2.Reaction (and S) EtOH AcO AcOH

alcohol-dehydrogenase

Name of the S + ase

(S-name)+reaction name + ase

3.Trivial names: + -in pepsine, tripsine, rennet or rennine protein degrading

4. IUB IUPAC 1964,1972,1978 Enzyme CommissionIUBMB

NOMENCLATURE OF ENZYMESTHEY ARE CALLED BY


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E.C.1.1.1.49. D-glucose-6P: NADP 1-oxydoreductase

Kind of reaction

Cosubstrate or coenzymeCatalog number

substrateLocation of attack is on the C1


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DataBank


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follows: ENZYMEKINETICS

Documents

1 Enzyme Intro