BABS2244 METABOLIC BIOCHEMISTRY

7/18/2019 BABS2244 METABOLIC BIOCHEMISTRY

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FACULTY OF APPLIED SCIENCES AND

COMPUTING (FASC)

KUALA LUMPUR CAMPUS

BACHELOR OF SCIENCE (HONS) IN

BIOSCIENCE WITH CHEMISTRY

BABS2244 METABOLIC BIOCHEMISTRY

EXPERIMENT 15

Red! E"#$%e&

NAME ID NO' PROGGROUP

Amirah Chan binti Latif 15WAR08520

RBS2 – Group1

Chey Sze Yin 15WAR0!1!"

Chon #hai $n 15WAR08%02

Goh Yee $n 15WAR08280



I"*d+,-" .

Bio&hemi&a' rea&tion( in 'i)in orani(m( are e((entia''y enery tran(fer(*

+n bio'oi&a' (y(tem(, o-i.ation i( often &oin&i.ent /ith the 'o(( of

hy.roen, .ehy.roenation* he enzyme( that &ata'yze the(e o-i.ation(

are &a''e. .ehy.roena(e(* -i.ationre.u&tion rea&tion( 3Re.o-rea&tion(4 mu(t o&&ur toether* $'e&tron( are tran(ferre. from the

re.u&in aent to the o-i.izin aent (u&h that the re.u&in aent i(

o-i.ize. an. the o-i.izin aent i( re.u&e.* +t i( &on)enient ho/e)er to

.e(&ribe the e'e&tron tran(fer rea&tion a( t/o ha'f rea&tion(, one for the

o-i.ation of the re.u&e. (pe&ie( an. one for the re.u&tion of the o-yen

(pe&ie(* Su&h &oup'e. rea&tion( are referre. to a( re.o- rea&tion(* he

metabo'i& pro&e((e( 'y&o'y(i(, #reb( Cy&'e, an. $'e&tron ran(port

6ho(phory'ation in)o')e the tran(fer of e'e&tron( 3at )aryin enery

(tate(4 by re.o- rea&tion(*

F-/+*e 1' 6a((ae of e'e&tron( from &ompoun. A to &ompoun. B*

3When A 'o(e( it( e'e&tron( it i( o-i.ize.7 /hen B ain( the e'e&tron( it i(

re.u&e.*4

F-/+*e 2' -i.ationre.u&tion )ia an interme.iary 3enery &arrier4

&ompoun., 9A:;*



$'e&tron( are tran(ferre. from one mo'e&u'e to another in four .i<erent

/ay(=

1 :ire&t'y a( e'e&tron(* 3>e2; ; Cu2; >e?; ; Cu;4

2 A( @y.roen atom(* he hy.roen atom &ontain( a proton, @; an.

an e'e&tron, e

A@2 A ; 2e ; 2@;

? A( a @y.ri.e ion, @* he hy.ri.e ion ha( t/o e'e&tron( an. i( hih'y

rea&ti)e* +n bio'oi&a' (y(tem( it i( .ire&t'y tran(ferre. to 9A:'ine.

.ehy.roena(e(*

" hrouh the .ire&t &ombination /ith o-yen* o'e&u'ar o-yen

&ombine( /ith orani& rea&tant( to o-i.ize hy.ro&arbon( to

a'&oho'(, a'.ehy.e( to a&i.(*

RDC@? ; E 2 RDC@2F@

A-% .

1* o .etermine the pre(en&e of enzyme( in )ariou( p'ant an. anima'

ti((ue(*

2* o &ompare the rate of enzyme a&ti)ity bet/een fre(h an. boi'e.(amp'e(*



DEHYDROGENASE ACTION

Me0d .

P* AA (erie( of 5 te(ttube( 'abe'e. A to $ /ere prepare.* 5 m' of 0*05

methy'ene b'ue (o'ution /a( a..e. in /ater to ea&h* he 'ean mea' /ith

eHua' (ma'' pie&e( that ha. prepare. by 'ab a((i(tant /a( treate. a(

in.i&ate. be'o/=

S%3e& C"e"&

e(ttube A 2 pie&e( of meat, fre(h'y &ut*

e(ttube B 2 pie&e( of meat, pre)iou('y boi'e. an. &oo'e.*

e(ttube C 2 pie&e( of meat, pre)iou('y min&e.*

e(ttube :2 pie&e. of meat, thorouh'y /a(he. un.er runnin tap/ater*

e(ttube $ Contro'*

After that, a'' 5 te(ttube( /ere &ore. an. 'eft un.i(turbe. in a /ater

bath maintaine. at a &on(tant temperature of ?%

C* he tube( /ere not(haen on&e the &ontent( ha. mi-e.* he &ontent( /ere ob(er)e. aain(t

a /hite ba&roun. an. any &hane( in the 5 tube( /ere re&or.e. at

inter)a'( of 5 minute(7 (tartin ", 5, 10, 15 Iminute( unti' t/o

(ub(eHuent ob(er)ation( (ho/ no &hane in &o'or, inten(ity an.

.i(tribution* he te(t tube( /ere un&ore. an. the &ontent( /ere (haen

thorouh'y* A'' ob(er)ation( an. the &on&'u(ion( /ere re&or.e. in a

tabu'ar form*

P* B



A .up'i&a a (erie( of ? te(ttube( &ontainin 5 m' of 0*05 methy'ene b'ue

/a( prepare. an. 'abe'e. them >a, >b, Ga, Gb, @a an. @b* he fo''o/in

fre(h (pe&imen( /ere a..e. to the JaK (erie( an. an eHua' Huantity of the

(ame (pe&imen pre)iou('y boi'e. an. &oo'e. /a( a..e. to the JbK (erie(

a( (ho/n in tab'e be'o/=

S%3e& C"e"&

e(ttube >2 pie&e( of 'i)er, fre(h'y &ut 3ea&h pie&e about 5mm&ube4

e(ttube G 2 pie&e( of potato fre(h'y &ut 3about 5mm &ube4

e(ttube @ 2 m' yea(t (u(pen(ion 310 yea(t (o'ution4*

A'' the ? pair( of te(ttube( /ere in&ubate. in a /ater bath at ?%C* Aain,

the &ontent( /ere ob(er)e. aain(t a /hite ba&roun. an. any &hane(

in the 5 tube( /ere re&or.e. at inter)a'( of 5 minute(7 (tartin ", 5, 10,

15 Iminute( unti' t/o (ub(eHuent ob(er)ation( (ho/ no &hane in

&o'our, inten(ity an. .i(tribution* he te(t tube( /ere un&ore. an. the

&ontent( /ere (haen thorouh'y* A'' ob(er)ation( an. the &on&'u(ion /e

ma.e /ere re&or.e. in tabu'ar form*

Re&+3& .



Observation at 5th minute










Observation at 35th minute (fnal)



P* A

S%3e& O&e*-"& Ded+,-"

e(ttube A So'ution &hane( from

b'ue to reeni(hb'ue*

6re(en&e of a

tran(parent 'ayer

aroun. the meat

:ehy.roena(e i( pre(ent*



e(ttube B So'ution remain( b'ue

ran(parent 'ayer at the

ba(e of te(t tube,

aroun. the meat i(

forme.

Some .ehy.roena(e i(

(ti'' pre(ent maybe

be&au(e meat /a( not

boi'e. proper'y*

e(ttube C So'ution &hane( from

b'ue to 'iht b'ue

:ehy.roena(e i( pre(ent,

but the amount i( 'e((

than te(ttube A*

e(ttube :

So'ution remain( b'ue

6 Greeni(hb'uetran(parent 'ayer at the

ba(e of te(t tube,

aroun. the meat i(

forme.

6Ab(en&e of

.ehy.roena(e*



e(ttube $ Co'or of (o'ution

remain( b'ue*

Ab(en&e of

.ehy.roena(e*



P* B

S%3e& O&e*-"& Ded+,-"

e(ttube >a So'ution &hane( from

b'ue to reeni(hb'ue*

So'ution i( ('iht'y

&'ou.y

:ehy.roena(e i( pre(ent*

e(ttube >b Co'or of (o'ution

remain( b'ue*

A ye''o/i(h 'ayer &an be

ob(er)e. aroun. theboi'e. 'i)er*

:ehy.roena(e enzyme(

are ('iht'y pre(ent*

he 'i)er /a( not boi'e.

proper'y*



e(ttube Ga Co'or of (o'ution

remain( b'ue*

:ehy.roena(e i(

ab(ent*

e(ttube Gb

6 Co'or of (o'ution

remain( b'ue*

6 :ehy.roena(e i(

ab(ent*

e(ttube @a So'ution turn( &'ou.y,

reeni(hb'ue &o'or

he yea(t i( (ta'e, thu( /e

&annot no/ that

/hether the yea(t&ontain( .ehy.roena(e

or not*



e(ttube @b So'ution turn( &'ou.y,

reeni(hb'ue &o'or

he yea(t i( (ta'e, thu( /e

&annot no/ that

/hether the yea(t

&ontain( .ehy.roena(e

or not*

D-&,+&&-" .

1' I" 70 7$& 0e 0e d-8e*e" *e%e"& " 0e %e8e,ed 0e-* ,-" " %e0$3e"e 3+e9 S+//e& &&-3e

e!3"-"&'

ne of the meat( i( bein boi'e. &omp'ete'y /hi'e the other meat i(

not* :ehy.roena(e( are pre(ent in meat(* hey initiate the o-i.ation

pro&e(( by tran(ferrin hy.roen from a (ub(trate to a hy.roen

a&&eptor* ethy'ene b'ue (o'ution i( often u(e. a( an arti&ia'

hy.roen a&&eptor* A &omp'ete'y boi'e. meat /i'' ha)e no

.ehy.roena(e( be&au(e they are .enature. un.er hih temperature*

hu(, no hy.roen i( pre(ent to be a&&epte. by the methy'ene b'ue



(o'ution an. it( &o'or remain( un&hane.* n the other han., an

unboi'e. meat ha( .ehy.roena(e(* @en&e, the .ehy.roena(e( /i''

tran(fer the hy.roen to the methy'ene b'ue (o'ution an. re.u&tion

o&&ur(* When re.u&e., methy'ene b'ue (o'ution /i'' &hane from b'ue

to &o'or'e((* he .e&o'orization of methy'ene b'ue (o'ution a&t( a( a)i(ib'e in.i&ator of the pre(en&e of .ehy.roena(e( in meat ti((ue* he

.eree of .ehy.roena(e( a&ti)ity a<e&t( the rate of .e&o'orization of

methy'ene b'ue (o'ution*

2' E!3-" 0e &-/"-:,",e ; &e-"/ + e&6+e E'

e(t – tube $ i( u(e. a( a &ontro' in the e-periment* here i( nothin to

be a..e. in(i.e thi( te(t – tube /hi&h mean( there /i'' be no rea&tion

in thi( te(t – tube* Be(i.e(, the &o'or of methy'ene b'ue in te(t – tube $

i( un&hane.* hu(, /e &an &ompare the &o'or of methy'ene b'ue in

te(t – tube( A to : /ith te(t – tube $ /hi&h it( methy'ene b'ue &o'or

/i'' ne)er bein &hane.* he &o'or &hane in te(t – tube( A to :

in.i&ate that there i( rea&tion in the te(t – tube(*

<' W0 -& 0e +*&e ; &e-"/ + +e& B= F= G "d H9

he(e tube( are bein (et up for &ompari(on of tube( A, >a, Ga an. @are(pe&ti)e'y* he ti((ue( u(e. in the(e te(t – tube( are bein

&omp'ete'y boi'e. an. &oo'e.* hu(, the hy.roena(e( /i'' be

.enature. an. there i( no rea&tion in(i.e the(e tube(* he &o'or of

methy'ene b'ue (o'ution /i'' remain un&hane. (in&e no hy.roen i(

pre(ent* So, /e &an ob(er)e /hether there i( rea&tion in the tube( A,

>a, Ga an. @a )ery &'ear'y by &omparin the &o'or of tube( B, >b, Gb

an. @b*

4' W0 de& &0>-"/ 0e ,"e"& -"d-,e + 0e *e,-" ;

0e *-+& %-!+*e&9

Shain the &ontent( i( important (o that the ti((ue( an. the (o'ution

&an mi- /e''* he &ontent( an. the (o'ution are bein mi-e. /e'' to

en(ure that there i( a &omp'ete rea&tion in the tube(* Ran.om error&an be a)oi.e. an. /e &an et more a&&urate ob(er)ation(*



5' P-" + &-%-3*--e& * d-8e*e",e& -" 0e e8e, ; 0e

d-8e*e" -&&+e& " %e0$3e"e 3+e'

>or a'' the boi'e. ti((ue( in thi( e-periment in&'u.in 'i)er, meat an.potato, there /i'' be no rea&tion /hen the(e boi'e. ti((ue( are bein

a..e. to the methy'ene b'ue (o'ution* hi( i( be&au(e methy'ene b'ue

(o'ution /hi&h i( the arti&ia' hy.roen a&&eptor &annot a&&ept

hy.roen /hi&h i( ab(ent in the(e boi'e. ti((ue(* Sin&e there i( no

hy.roen to be re&ei)e. by methy'ene b'ue (o'ution, the methy'ene

b'ue (o'ution /i'' remain un&hane. /hi&h i( b'ue &o'or* n the other

han., for the (ame ti((ue( (u&h a( boi'e. meat an. unboi'e. meat,

they /i'' be put into the te(t tube( /ith (ame (o'ution* After 25 – ?0

minute(, /e &an ob(er)e that the in te(t tube of boi'e. meat, themethy'ene b'ue (o'ution turn( to &o'or'e(( but in the te(t tube of

unboi'e. meat, the methy'ene b'ue (o'ution remain( b'ue* hi(

in.i&ate( that the hy.roena(e( in boi'e. meat are .enature. /hi'e

the hy.roena(e( in unboi'e. meat are not bein .enature.*

?' W0 ,**e3-" ," $+ d*7 + 0e ,--$ "d 0e

&+*,e ; 0e e"#$%e9

:i<erent (our&e /i'' &ontain .i<erent amount of enzyme* he enzyme

/i'' (pee. up the rea&tion by 'o/er the a&ti)ation enery* he rate of

the rea&tion .epen.( on the pre(en&e of enzyme* +f there are more

enzyme(, the rea&tion /i'' be fa(ter &ompare. to the rea&tion /ith

fe/er enzyme(*

CATALASE ACTION

Me0d .



A .up'i&ate (erie( of 5 te(ttube( 'abe'e. Aa, Ab, Ba, BbI$a, $b /ere

prepare.* hen, 5 m' of hy.roen pero-i.e 310 )o'ume(4 /a( a..e. to

ea&h te(t tube* he fo''o/in fre(h (pe&imen( a( (ho/n in tab'e be'o/

/ere a..e. to the JaK (erie( an. a (imi'ar Huantity of the (ame (pe&imen(

pre)iou('y boi'e. for 5 minute( an. &oo'e. /a( a..e. to the JbK (erie(=

S%3e& C"e"&

e(ttube A 2 pie&e( of meat 3ea&h 5mm &ube4*

e(ttube B 2 pie&e( of 'i)er 3ea&h 5mm &ube4*

e(ttube C 2 pie&e( of potato 3ea&h 5mm &ube4*

e(ttube : 2 pie&e( of app'e 3ea&h 5mm &ube4*

e(ttube $ 2 m' of yea(t (u(pen(ion*

e(ttube >A pie&e of p'ant (tem 'eaf &ut out of the entire oran/ith a (harp b'a.e (o that it pre(ent( both the inta&t(urfa&e an. &ut &e''(*

e(ttube GA (amp'e of potato meat p'u( !5 a'&oho' to &o)er*A''o/ to (tan. for 5 minute(* 6our out the a'&oho'*

e(ttube @ A pin&h of manane(e .io-i.e*

he freHuen&y an. (ize of e(&apin bubb'e( /ere ob(er)e. for ea&h te(t

tube* he ob(er)ation( an. &on&'u(ion( for ea&h te(t tube /ere tabu'ate.*

Re&+3& .

S%3e& O&e*-"& Ded+,-"



e(ttube Aa $<er)e(&en&e &ou'. be

ob(er)e. /hen the

meat i( .roppe. in*

Cata'a(e i( pre(ent*

e(ttube Ab $<er)e(&en&e o&&ur(

('o/'y /hen boi'e. meat

/a( .roppe. in

Cata'a(e i( pre(ent in

(ma'' Huantity*

e(ttube Ba $<er)e(&en&e o&&ur(

)ery rapi.'y /hen the

'i)er /a( .roppe. in


'are amount*



e(ttube Bb 9o bubb'e( forme. Cata'a(e i( ab(ent

be&au(e the 'i)er ha.

been boi'e.*

e(ttube Ca 6 >e/ noti&eab'e bubb'e(

forme.*


(ma'' Huantity*

e(ttube Cb 6 >e/ noti&eab'e bubb'e(

forme.*


(ma'' Huantity*



e(ttube :a6 Mery fe/ bubb'e(

forme.* Cata'a(e i( pre(ent in

(ma'' Huantity*

e(ttube :b Mery fe/ bubb'e(

forme.*


(ma'' Huantity*

e(ttube $a6 9o e<er)e(&en&e

o&&urre.*


yea(t, but in thi( &a(e, the

yea(t i( (ta'e, thu( the

re(u't /a( a<e&te.*



e(ttube $b 9o e<er)e(&en&e

o&&urre.*


yea(t, but in thi( &a(e, the

yea(t i( (ta'e, thu( the

re(u't /a( a<e&te.*

e(ttube > Bubb'e( forme. aroun.

the e.e( of the 'eaf*

Cata'a(e i( pre(ent in the

p'ant ti((ue(*

e(ttube G $<er)e(&en&e o&&ur(

rapi.'y*

Cata'a(e i( pre(ent*



e(ttube @ Mery 'itt'e e<er)e(&en&e* Cata'a(e i( pre(ent*

anane(e .io-i.e i( an

inorani& &ata'y(t 3but not

an enzyme4 for thebreain .o/n of

hy.roen pero-i.e*

D-&,+&&-" .

1' W0 -& 0e e8e, ; -3-"/ " 0e d-8e*e" 3" "d "-%3

-&&+e&9

ne of the fa&tor( that &au(e( enzyme to be .e(troye. i( the e-po(ure

of the enzyme( to heat* When boi'in i( &arrie. out, e-&e((i)e heat i((upp'ie. to the p'ant an. anima' ti((ue(, &au(in the &e''( to be i''e.

an. enzyme( are e)entua''y .enature.*

2' W0$ -& - "e,e&&*$ &e + 0e @ &e*-e& ; e& +e&9

he JbK (erie( of te(t tube( a''o/( u( to ob(er)e /hether &ata'a(e in

the(e boi'e. ti((ue( i( ab'e to &ata'yze the .e&ompo(ition of hy.roen

pero-i.e* herefore /e &an &ompare it( rate of enzyme a&ti)ity /ith it(re(pe&ti)e fre(h ti((ue(*

<' De& -3-"/ 8e, 0e ,-" ; %"/"e&e d-!-de9 W0$9

Boi'in i( ab'e to a<e&t the a&tion of manane(e .io-i.e, but the

e<e&t( are not too noti&eab'e* hi( i( be&au(e manane(e .io-i.e i( an

inorani& &ata'y(t an. inorani& &ata'y(t( are 'e(( (en(iti)e to hih

temperature(* herefore, e<er)e(&en&e &ou'. be ob(er)e. in te(ttubeG*



4' I& 0e*e "$ d-8e*e",e -" 0e %+" ; /& e3ed 70e"

d-8e*e" -&&+e& *e +&ed9 W0 ,",3+&-"& ," $+ d*7

*e/*d-"/ 0e ,--$ ; 0e e"#$%e 70e" - -& ; d-8e*e"

-&&+e *-/-"9

Ba(e. on the ob(er)ation(, more a( i( e)o')e. /hen an anima' ti((ue

i( u(e. &ompare. to p'ant ti((ue(, /hereby the fre(h 'i)er (ho/( the

hihe(t amount of a( e)o')e.* Cata'a(e, /hi&h &ata'yze( the

.e&ompo(ition of hy.roen pero-i.e into o-yen an. /ater 3re(u'tin

in an e<er)e(&en&e4, e-hibit( a hiher a&ti)ity in anima' ti((ue( a(

anima'( ha)e a reater nee. for metabo'i(m an. &e''u'ar re(piration* +t

fun&tion( a( a prote&ti)e enzyme an. a''o/( 'i)in orani(m( to eep

'i)in, /hi&h i( &ru&ia' e(pe&ia''y in anima'(*

C",3+&-" .

@y.roena(e( are &ommon'y pre(ent in p'ant an. anima'( ti((ue(* +n thi(

e-periment, it( fun&tion i( to tran(fer hy.roen from the (ub(trate /hi&h

i( the ti((ue( to the hy.roen a&&eptor /hi&h i( the methy'ene b'ue

(o'ution* he methy'ene b'ue (o'ution /i'' be re.u&e. /hen hy.roen i(

bein a&&epte. an. the &o'or /i'' &hane from b'ue to &o'or* he &o'or

&hane of methy'ene b'ue (o'ution in.i&ate( that there i( rea&tion*

Be(i.e(, the rate of enzyme a&ti)ity for fre(h ti((ue( i( fa(ter than the

rate of enzyme a&ti)ity for boi'e. ti((ue(* hi( i( .ue to the hy.roena(e(

in boi'e. ti((ue( i( .enature. un.er hih temperature /hi'e hy.roena(e(in fre(h ti((ue( i( (uN&ient for rea&tion to tae p'a&e* hu(, the rate of

enzyme a&ti)ity for boi'e. ti((ue( /i'' be ('o/er*

Re;e*e",e& .

1* Ra(toi, MB 1!!%, Modern biology, 6itambar 6ub'i(hin Company 364

Lt.*, 9e/ :e'hi*



2* :i<en 2015, Catalyst vs enzyme, )ie/e. 8 Oune 2015,

Phttp=///*.i<en*

&om.i<eren&eCata'y(tQ)(Q$nzyme

?* a.S&i 9et/or 1!!8, e! Catalase a"tivity in various "ells, )ie/e. 8

Oune 2015, Phttp=///*ma.(&i*orpo(t(ar&hi)e(1!!8

11!1088518%*B&*r*htm'

"* Cata'a(e by Oennifer &:o/a'', )ie/e. on 12th Oune 2015,

Phttp=///*ebi*a&*u

interpropotm200"Q!6ae1*htm

5* ethy'ene B'ue Re.u&ta(e e(t, )ie/e. on 12th Oune 2015,

Phttp=)'ab*amrita

*e.u(ubT?Ubr&hT%?U(imT1V?0U&ntT1

BABS2244 METABOLIC BIOCHEMISTRY

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