Biochem. Exp2

8/17/2019 Biochem. Exp2

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Bio-ChemicalEngineering

Experiment.2 Enzyme

Name: Kareem Tarek

I.D: 115780

Chemical- Year 3

Supervised by:

• Dr. Mohamed Dessouki

• TA. mar A!o E" Azayem


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Contentso A!stra#t............................................................................2

o $ntrodu#tion.......................................................................2

o Experimenta" Methods and %ro#edure...............................&

o Experimenta"

'esu"ts……………………………………………………………………..5o Dis#ussion.........................................................................8

o (our#es o) Error.................................................................8

o 'e)eren#es........................................................................*

List of i!ures+i,ure 1 -o#k Key Me#hanism/ ompetitie $nhi!ition............................................

+i,ure 2 3on/ ompetitie $nhi!ition..........................................................................

+i,ure E4e#t o) temperature on rea#tion rate..........................................................&

+i,ure & E4e#t o) p on rea#tion rate.........................................................................5

+i,ure 5 Enzyme on#entration s. Enzyme A#tiity................................................10

1 | % a , e


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A!stra#t The o!6e#tie o) this experiment is to inesti,ate and ,ain kno"ed,e o) the

physi#a" and #hemi#a" aspe#ts o) Enzymes and to #ondu#t seera" tests to

eri)y and o!sere e4e#ts o) on#entration Temperature and $nhi!itors on

enzymes. 9y #ondu#tin, these tests students i"" !e a!"e to determine the

optimum #onditions )or enzyme a#tiity.

$ntrodu#tionEnzymes are protein mo"e#u"es in nature that sere as a !io"o,i#a" #ata"yst.

Enzymes are )ormed !y "iin, #e""s su#h as p"ants anima"s and mi#roor,anisms.

They are un:uestiona!"y #ru#ia" as #ata"ysts in !io#hemi#a" rea#tions. %ra#ti#a""y

eery rea#tion in a #e"" ino"es the mani)estation o) a spe#i;# enzyme. ne o) the

ma6or )un#tions o) enzymes in a "iin, system is to #ata"yze the makin, and

!reakin, o) #hemi#a" !onds. onse:uent"y "ike any other #ata"ysts they in#rease

the rate o) rea#tion ithout themse"es under,oin, permanent #hemi#a" #han,es.

The #ata"yti# a!i"ity o) enzymes is due to its parti#u"ar protein stru#ture.

$. An enzyme #ata"yst is ex#eedin,"y spe#i;# and #ata"yzes on"y one or a sma""

num!er o) #hemi#a" rea#tions. A ast assortment o) enzymes exist hi#h #an

#ata"yze an ex#eptiona""y #omprehensie ran,e o) rea#tions.$$. The rate o) an enzyme/#ata"yzed rea#tion is #ustomari"y mu#h )aster than

that o) the same rea#tion hen dire#ted !y non/!io"o,i#a" #ata"ysts. n"y a

tri?in, amount o) enzyme is ne#essary to produ#e a desired e4e#t.$$$. The rea#tion #onditions


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% &odulator


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i!ure + Non- Competitive Inhibition

",ect of emperature on "n#ymes

The rate o) a #hemi#a" rea#tion is dependent on the temperature #orrespondin, to

Arrhenius e:uation as>

(u!se:uent"y hen -n is p"otted s. 1I a strai,ht "ine ith s"ope -"I$ is o!tained.

The temperature dependen#y o) numerous enzyme/#ata"yzed rea#tions #an !e

termed !y the Arrhenius e:uation. A rise in the temperature upsur,es the rate o)

rea#tion sin#e the atoms in the enzyme mo"e#u"e hae ,rander ener,ies and an

immense in#"ination to moe. 3eerthe"ess the temperature is restri#ted to the

standard !io"o,i#a" ran,e.


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i!ure 3 ",ect of temperature on reaction rate

",ect of p/ on "n#ymes The rate o) an enzyme rea#tion is intense"y in?uen#ed !y the p o) the rea#tion

so"ution !oth in vivo and in vitro. The optimum p aries )rom one enzyme to the

other. +or instan#e pepsin )rom the stoma#h has an optimum p !eteen 2 and.

hi"e the optimum p o) amy"ase )rom sa"ia is .8.hymotrypsin )rom the

pan#reas has an optimum p in the mi"d"y a"ka"ine re,ion !eteen 7 and 8.

• Enzyme is a protein hi#h #omprises o) ammo a#id ;"trates


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• The amino a#id residues possess !asi# neutra" or a#id side ,roups hi#h #an

!e positie"y or ne,atie"y #har,ed at any ,ien p.

i!ure 0 ",ect of p/ on reaction rate

Experimenta" Methods and %ro#edure

est based on the of "n#yme Concentration

Materia"s>1 (tar#h (o"ution

ommer#ia" amy"ase so"ution

$odine rea,ent

6 | % a , e


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9enedi#tFs rea,ent

%ro#edure>1. -a!e" ;e test tu!es 1/5. %"a#e & m- o) 1 star#h in ea#h o) the ;rst )our test

tu!es. %"a#e & m- o) amy"ase so"ution in the ;)th tu!e. %"a#e a"" o) the tu!es in the

7C ater !ath )or 5 minutes. !tain 5 #"ean droppers and "a!e" them 1/5.


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2/ %"a#e a star#h tu!e and an amy"ase tu!e in the 7C ater !ath. %"a#e one tu!e

o) ea#h in an i#e/ater !ath and one o) ea#h tu!e in a !oi"in, ater !ath. Keep the

tu!es in their !aths )or 10 minutes to a""o them to rea#h the temperature o) their

!aths.

/ 'ead and re#ord the temperature o) the i#e/ater !ath. %our the amy"ase so"ution

into the star#h so"ution mix and put the tu!e !a#k into the !ath. 'e#ord the time.

&/ 'epeat step )or the 7C !ath.

5/ 'epeat step )or the !oi"in, ater !ath.

/ +or ea#h mixture a)ter 10 minutes o) rea#tion hae e"apsed trans)er & drops

o)the mixture to a spot p"ate. Add 1 drop o) iodine rea,ent to ea#h samp"e. 'e#ord

the #o"or and a#tiity "ee" o) the enzyme in ea#h #ase.

7/ raph the enzyme a#tiity "ee" s. temperature

est based on the e,ect of Inhibitors on en#ymes

Materia"s>1 (tar#h so"ution ommer#ia" amy"ase so"ution

1 3a" *5 ethano"

A,3I $odine rea,ent

%ro#edure>1/ %"a#e & m- o) amy"ase so"ution in ea#h o) three test tu!es. To the ;rst tu!e add

10 drops o) 1 3a" so"ution. Add 10 drops o) *5 ethano" to the se#ond test

tu!e. Add 10 drops o) either A,3 or %!


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Experimenta" 'esu"ts

%art .1J (amp"e. 1"n#yme concentration12 molmL

Time (minutes) Observation (color) Enzyme activity 0.5 9"ue/!"a#k 0


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5 o"den !ron 2


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Dis#ussion

The resu"ts a!oe sho the temperature e4e#ts on the a#tiity o) enzymes. 9ased

on the experimenta" resu"ts it as #on#"uded that the optimum a#tiity )or enzymes

ere at 7C the amy"ase in?uen#ed the star#h samp"e !y a"terin, its #o"or to

!ron. At 0C the amy"ase a#tiity diminished !y a"terin, the #o"or to dark !ron

hi#h in)ers that it the e4e#t o) amy"ase has a"tered the properties o) the star#h. At

100C the amy"aseHs a#tiity as exhausted it yie"ded a !"ue/!"a#k #o"or hi#h

indi#ates that no #han,es ere made to the properties o) star#h.

(our#es o) Error To aoid any errors that i"" render ina##urate resu"ts it is adised to>

A. To thorou,h"y ash the test tu!es ith disti""ed ater and remoe any residue

so"utions !e)ore #ondu#tin, the experiment.

9. -a!e" eery test tu!e a##ordin, to the so"ution it ho"ds to aoid mixin, the

samp"es.

. Gse a stopat#h to proper"y time the heat re:uired )or a samp"e to rea#t.

D. (tudents o)ten add the ron, rea,ent to the samp"e it is adised that ea#hstudent shou"d hand"e one samp"e to aoid addin, the ron, rea,ent.

'e)eren#es

• Dutta '.

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