enzim laporan praktikum biokimia

14
LAPORAN PRAKTIKUM BIOKIMIA DASAR “KINETIKA ENZIM” DISUSUN OLEH :  NAMA : JUNIFER TULUNGEN  NIM : 13 533 030 KELAS : C KELOMPOK : 1 (SAT U) JURUSAN KIMIA FAKULTAS MATEMATIKA DAN ILMU PENGETAHUAN ALAM UNIVERSI TA S NEGERI MANADO NOVEMBER 2015

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Page 1: Enzim Laporan Praktikum Biokimia

8/18/2019 Enzim Laporan Praktikum Biokimia

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8/18/2019 Enzim Laporan Praktikum Biokimia

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A. Tuju!

Perco!!" #"# $#r!"c!"% &"'& e*ere"!+!" 'e"'!"% e",# $!" -!'or.

-!'or /!"% e*e"%!r&# !'##'!2"/! E",# /!"% !!" #'! *e+!4!r# !$!+!

e",# !'!+!2e /!#'& e",# /!"% er-&"%2# eec! #$ro%e" *ero2#$! e"4!$#

!#r $!" %!2 o2#%e"

B. D"# T$%#&

1 E",#

D!+! 2/2'e #o+o%# re!2# ##! 2e+!+& eer+&!" !'!+#2 T!"*!

!'!+#2 re!2# ##! er+!"%2&"% 2!"%!' +!! 2e#%%! $#*er+&!" e",#

E",# er-&"%2# 2e!%!# #o!'!+#2!'or E",# er&*!!" 2!+! 2!'& ,!'

 #o+o%#2 /!"% *!+#"% *e"'#"% /!"% 'er$!*!' *!$! or%!"#2 #$&* (T# Do2e"

Pr!'#& 6#o##! D!2!r 7015)

E",# !$!+! o+e&+ #o*o+/er /!"% 'er2&2&" !'!2 !2! !#"o

$!+! o*o2#2# $!" 2&2&"!" r!"'!# 'er!'&r $!" 'e'!* Se!%!# *ro'e#" e",#

$#*ro$&2# $!" $#%&"!!" o+e 2e+ #$&* &"'& e"%!'!+#2#2 re!2# !"'!r!+!#" o"er2# e"er%/ $!" e'!o+#2 *er'!!"!" 2e+ E",# $#%o+o"%!"

e"&r&' re!2# /!"% $##&'#"/! 2e$!"%!" "!! e",# $#er#!" "!!

e"&r&' 2&2'r!'"/!(A+$#7010)

T!'! "!! e",# /!"% !r& #!2!"/! er!2!+ $!r# !' "u(")#)!* $e"%!"

e"!!!" !#r!" +"$ Se!%!# co"'o 2e&! e",# /!"% eer4! *!$!

+#*#$ !!" $#2e&' +#*!2e 6er#&' er&*!!" 2&2&"!" 2e$er!"! $!r# %!&"%!"

!"'!r! e",# $!" 2&2'r!':

e",# 8 2&2'r!' o*+e2 e",#.2&2'r!' e",# 8 *ro$& 

E + S ↔ ES ↔ EP ↔ E + P

Se'e+! *ro$& $#+e*!2!" e",# e&$#!" e!2 &"'& e"%#!' o+e&+

2&2'r!' +!#" $!" *ro2e2 $#!'!2 $#&+!"% e!+# J&+! o+e&+ 2&2'r!' /!"%

$!*!' 'er#!' $!" ere!2# *er e"#' $#2e&' "#+!# turnover  E",# !'!+!2e /!"%

e"%&r!# #$ro%e" *ero2#$! e"4!$# !#r $!" %!2 o2#%e" e#+## "#+!#

turnover 2ee2!r +#! 4&'! I"# er!r'# !9! $!+! 2!'& e"#' 2e'#!* o+e&+

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e",# !*& eec! +#! 4&'! o+e&+ #$ro%e" *ero2#$! (T# Do2e"

Pr!'#& 6#o##! D!2!r 7015)

E",# $#!%# $!+! e"! %o+o"%!" e2!r o+e Co##2#o" o" E",/e2 o- 

'e I"'er"!'#o"!+ U"#o" o- 6#oce#2'r/Pe"%%o+o"%!" #"# $#$!2!r!" !'!2

re!2# ##! $#!"! e",# ee%!"% *er!"!" *e"'#"% D!+! ##! $#e"!+

 eer!*! #2'#+! *e"'#"% /!#'&

• Koe",# !'!& o-!'or /!"% 'er#!' 2!"%!' &!' !!" 'er#!' $e"%!"

#!'!" o!+e" $e"%!" e",# à  gugus prostetik  

• E",# !'#- +e"%!* $e"%!" 2e&! o*o"e""/! (%&%&2 *ro'e#" !&*&"

"o" *ro'e#")à holoenzim 

• 6!%#!" /!"% 'er$#r# $!r# *ro'e#" 2!4! *!$! 2&!'& e",# à  Apoenzim  ;

apoprotein 

• F&"%2# oe",# !$!+! 2e!%!# !r#er 2ee"'!r! $!r# %&%&2 -&"%2#o"!+ /%

 er*er!" $!+! re!2# e"2#!'#2 'er2e&'(Poe$4#!$#700<)

E",# !!" e#+!"%!" !'#!2#"/! !re"! *!"!2 !2! $!" !2! &!'*e+!r&'

or%!"#c !'!& !*! 2!4! /!"% e"/e!!" $e"!'&r!2# *ro'e#" (G#r#"$!1=><)

Mec!"#2 o- E",#e !c'#/'#

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2. F,)%#+-,)%# *! '$'/$!#u& ,)&&)" $!&'

! Ko"2e"'r!2# 2&2'r!'

  Ko"2e"'r!2# E",#

c *H ('#"%!' e!2!!" )

$ S&&

e A$!"/! o-!'or 

- A$!"/! #"##'or 

3. S&"& ,)&- 

S#2# !'#- e*&"/!# 7 !%#!" /% *e"'#"%:

o 6!%#!" /!"% e"%e"!+ 2&2'r!' $!" e&$#!" e"%#!'"/!

o 6!%#!" /!"% e"%!'!+#2#2 re!2# 2e'e+! 2&2'r!' $##!' o+e e",#

A2! !#"o /!"% ee"'& e$&! !%#!" 'er2e&' '#$! !r&2 er$e!'!"

$!+! &r&'!" 2ec!r! +#"e!r 'e'!*# $!+! o"-or!2# 3D ere! er$e!'!"

4. T$%#& u!)u, '$!j$",! ,$#j $!&'

 Lock and Key analogy

E",# e#+## 2'r&'&r 2#2# 2*e2#-# /!"% coco $e"%!" 2&2'r!'

M!*& e"er!"%!" 2*e2#-#'!2 e"2# ''* '#$! $!*!' e"er!"%!" 2'!#+#'!2

-!2e 'r!"2#2# e"2#

 Induced Fit theory

e*er'#!"%!" -+e2##+#'!2 *ro'e#" 2e#"%%! *e"%#!'!" 2&!'& 2&2'r!'

 *!$! e",# e"/e!!" 2#2# !'#- e"%&! o"-or!2#"/! 2e#"%%! coco $%"

2&2'r!'"/! à $*' e"er!"%!" -!2e 'r!"2#2# ES o*+e  

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> U&r '#"%%# %e+e&"%/!"% 'ere"'& (c) *!$! '#!* '!&"% 2e'e+! 1

e"#' 2er'! #'&"%+! o+&e %e+e&"% /!"% 'ere"'& (c3 !'!& L)

$!" c!'!'+! !2#+ *e"%#'&"%!""/! *!$! '!e+ !2#+ *e"%!!'!"

Ge+e&"% /!"% 'ere"'& e"!"$!!" $#!2#+!""/! %!2 o2#%e" (O7)

= U+!"%# *ro2e$&r "o ? > &"'& '!&"% 7 $!" e&$#!" '!&"% 3

10 H#'&"%+! +!4& *ee"'&!" O7(c3;2 !'!& L;2) ($##'&"% $!r# o+&e

%e+e&"% (c3 !'!& L) $#!%# $e"%!" 9!'& (1 e"#' B <0 2)

11 6&!'+! %r!-# %!r#2 /!"% e"&"4&!" &&"%!" !"'!r! +!4&

 *ee"'&!"O7(2&& /) $e"%!" *H +!r&'!" (2&& )

2.P$!#u ,%!"$!)#"& $!&' )$#7/ ,)&&)" $!&' ,)"$

  A) 7! B! 9

T!&"% re!2# 8 r! '!&"% *e"%%!r#2 %e+!2 &&r 10 L %e+!2 *#!+!;e!er 

750 L *#*e' 'e'e2 *#*e' o+&e'r# 5 L or'!r 8 !+& 2*!'&+!  stopwatch

!rer;+!e+ *#2!& '!+e"!" grater ;+e"$er '#!"%!" e"'!"% +!r&'!" H7O7

!&!$e2

 

6# ,$#j 9

1 6&!'+! e2'r! e"'!"% $e"%!" e"#!"% 2e#'!r 100 %r! e"'!"%

/!"% 'e+! $#&*!2 $#c&c# er2# $!" $#*o'o"%.*o'o"% ec#+ $!" $#+e"$er 

 er2!! $e"%!" !&!$e2 2e#'!r 100 ? 700 L

7 E2'r! e"'!"% $#'!*&"% $!+! %e+!2 *#!+!;e!er 750 L (E2'r! 

e"'!"% /!"% $#*ero+e $!*!' $#%&"!!" *!$! *erco!!" 2e+!"4&'"/!)

3 D#&!' !r#!2# o"2e"'r!2# e",# !'!+!2e /!"% 'er$!*!' $!+! e2'r! 

e"'!"% $!" $#re!2#!" $e"%!" +!r&'!" H7O7 $!" !&!$e2 $!+! '!&"%

re!2# $e"%!" o*o2#2# 2e!%!# er#&' :

! 05 L e2'r! e"'!"% (+!r&'!" e",#) $#c!*&r $e"%!" 3 L +!r&'!"

H7O7 (2&2'r!') $!" 75 L !&!$e2 $!+! '!&"% 1

  1 L e2'r! e"'!"% (+!r&'!" e",#) $#c!*&r $e"%!" 3 L +!r&'!"

H7O7 (2&2'r!') $!" 7 L !&!$e2 $!+! '!&"% 7

c 15 L e2'r! e"'!"% (+!r&'!" e",#) $#c!*&r $e"%!" 3 L +!r&'!"

H7O7(2&2'r!') $!" 15 L !&!$e2 $!+! '!&"% 3$ 7 M+ e2'r! e"'!"% (+!r&'!" e",#) $#c!*&r $e"%!" 3 L +!r&'!"

H7O7(2&2'r!') $!" 1 L !&!$e2 $!+! '!&"% 3

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@ A!'# re!2# /!"% 'er4!$# U&r+! '#"%%# %e+e&"% /!"% &"c&+(c)

 *!$! '#!* '!&"% (1 ? 3) 2e'e+! 1 e"#' #'&"%+! o+&e %e+e&"%

/!"% $#!2#+!" (c3 !'!& L) $!" c!'!' !2#+ *e"%#'&"%!" $!+! '!e+

!2#+ *e"%!!'!"

5 H#'&"%+! +!4& *ee"'&!" O7 (c3;2 !'!& L;2)

< 6&!'+! %r!-# %!r#2 /!"% e"&"4&!" &&"%!" !"'!r! +!4&

 *ee"'&!"O7(2&& /) $e"%!" o"2e"'r!2# e",#(2&& )

3.P$!#u ,%!"$!)#"& "u(")#) )$#7/ ,)&&)" $!&' ,)"$

 

6# ,$#j 9

1 D#&!' !r#!2# o"2e"'r!2# 2&2'r!' (+!r&'!" H7O7) $!" $#re!2#!" $e"%!"

e2'r! e"'!"% (+!r&'!" e",#) $!" !&!$e2 $!+! '!&"% re!2# $e"%!"

o*o2#2# 2e!%!# er#&' :

! 3 L e2'r! e"'!"% (+!r&'!" e",#) $#c!*&r $e"%!" 1 L +!r&'!" H7O7

(2&2'r!') $!" 7 L !&!$e2 $!+! '!&"% 1

  3 L e2'r! e"'!"% (+!r&'!" e",#) $#c!*&r $e"%!" 15 L +!r&'!"

H7O7 (2&2'r!') $!" 15 L !&!$e2 $!+! '!&"% 7

c 3 L e2'r! e"'!"% (+!r&'!" e",#) $#c!*&r $e"%!" 7 L +!r&'!"

H7O7 (2&2'r!') $!" 1 L !&!$e2 $!+! '!&"% 3

$ 3 L e2'r! e"'!"% (+!r&'!" !'!+!2e) $#c!*&r $e"%!" 75 L +!r&'!"

H7O7 (2&2'r!') $!" 05 L !&!$e2 $!+! '!&"% @

e 3 L e2'r! e"'!"% (e",# !'!+!2e) $#c!*&r $e"%!" 3 L +!r&'!"

H7O7 (2&2'r!') $!" 00 L !&!$e2 $!+! '!&"% 5

7 A!'# re!2# /!"% 'er4!$# U&r+! '#"%%# %e+e&"% /!"% &"c&+ (c) *!$! '#!* '!&"% (1 ? 5) 2e'e+! 1 e"#' #'&"%+! o+&e %e+e&"%

/!"% $#!2#+!" (c3 !'!& L) $!" c!'!' !2#+ *e"%#'&"%!" $!+! '!e+

!2#+ *e"%!!'!"

3 H#'&"%+! +!4& *ee"'&!" O7 (c3;2 !'!& L;2)

@ 6&!'+! %r!-# %!r#2 /!"% e"&"4&!" &&"%!" !"'!r! +!4&

 *ee"'&!"O7(2&& /) $e"%!" o"2e"'r!2# 2&2'r!'(2&& )

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D. H"& P$!')!

1. Pengaruh perubahan pH terhadap aktivitas enzim katalase

 No

T!&"%

H!2#+ Pe"%!!'!"

 *H

+!r&'!"

T#"%%#

Ge+e&"%

O7 (c)

o+&e

Ge+e&"%

O7(c3 !'!&

L)

L!4&

Pee"'&!"

O7 (c3;2 !'!&

L;2)1 1 (A2!) 3 05 03= 00<

7 7 (6!2!) 11 @ 31@ 005

3 3 (Ne'r!+) 5 0> 0<7 001

U"'& o+&e %e+e&"% U"'& +!4& *ee"'&!"

1   V =π r2

.t  1 BV 

B 31@ 057 05 B0,3960

B 31@ 075 05 B00<

B 03=

7   V =π r2

.t  7 BV 

B 31@ 057  @ B3,14

60

B 31@ 075 @ B 005

B 31@

3   V =π r2

.t  3 BV 

B 31@ 057 0> B3,14

60

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B 0<7 B001

2. Pengaruh konsentrasi enzim terhadap aktivitas enzim katalase

 No

T!&"%

H!2#+ Pe"%!!'!"

L!r&'!"

e",#

(L)

T#"%%I

Ge+e&"%

O7 (c)

o+&e

Ge+e&"% O7

(c3 !'!& L)

L!4&

Pee"'&!"

O7  (c3;2 !'!&

L;2)

1 1 05 0< 0@ 000>

7 7 1 10 0> 0013

3 3 15 1< 175 0071

@ @ 7 05 03= 000<

U"'& o+&e %e+e&"% U"'& +!4& *ee"'&!"

1   V =π r2

.t  1 BV 

B 31@ 057 0< B0,47

60

B 31@ 075 0< B000=

B 0@

7   V =π r2

.t    7 BV 

B 31@ 057  10 B0,78

60

B 31@ 075 10 B 001

B 0>

3   V =π r2

.t    3 BV 

B 31@ 057  1< B1,25

60

B 175 B007

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@   V =π r2

.t    3 BV 

B 31@ 057  05 B0,39

60

B 03= B000<

3. Pengaruh konsentrasi substrat terhadap aktivitas enzim

katalase

 No

T!&"%

H!2#+ Pe"%!!'!"

L!r&'!"

H7O7

(L)

T#"%%#

Ge+e&"%

O7 (c)

o+&e

Ge+e&"% O7

(c3 !'!& L)

L!4&

Pee"'&!"

O7  (c3;2 !'!&

L;2)

1 1 10 7> 71= 003

7 7 15 3 735 0033 3 70 5 3=7 00<

@ @ 75 55 @31 00

5 5 30 < @1 011

U"'& o+&e %e+e&"% U"'& +!4& *ee"'&!"

1   V =π r2

.t  1 BV 

B 31@ 057 7> B

2,91

60

B 31@ 075 7> B 003

B 0@

7   V =π r2

.t    7 BV 

B 31@ 057  3 B 2,3560

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B 31@ 075 3 B 003

B 0>

3   V =π r2

.t    3 BV 

B 31@ 057  5 B3,92

60

B 175 B00<

@   V =

π r

2

.t    @ B

B 31@ 057  55 B4,31

60

B 03= B 00

5   V =π r2

.t    5 BV 

B 31@ 057  < B4,71

60

B 175 B011

E Pe!!2!"

Pr!'#& /!"% !# +!&!" !+# #"# /!#'& 'e"'!"% e",#/!"%+e# 2*e2#-# e"%!"!+#2#2 'e"'!"% e",# !'!+!2e /!"% 'er$!*!' *!$!

e2'r! e"'!"% E",# !$!+! *ro'e#" /!"% er*er!" 2e!%!# !'!+#2 $!+!

e'!o+#2 !+& #$&* E",# er*er!" &"'& e*erce*!' re!2# ##!

/!"% 'er4!$# $!+! '&& !+& #$&* 'e'!*# e",# #'& 2e"$#r# '#$! #&'

 ere!2#

E",# !'!+!2e er&*!!" 2!+! 2!'& e",# /!"% 'er$!*!' *!$!

'&&!" E",# $#*ro$&2# o+e *ero2#2o $!" !'#- e+!&!" re!2#

o2#$!'#- !!".!!" /!"% $#!"%%!* 'o2# o+e '!"!!" 2e*er'# /$ro%e"

 *ero2#$! (H7O7)

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P!$! *r!'#& #"# /!#'& e"%e"!# e",# !'!+!2e $e"%!" '&4&!"

/!#'& &"'& eec! /$ro%e" *ero2#$! e"4!$# !#r $!" %!2 o2#%e"

$!+! *r!'#& #"# 'er$!*!' 5 *ro2e$&r *erco!!" "!&" !re"! eer!*!

!+ !! /!"% $#+!&!" !"/! 3 *erco!!" 2!4! e'#%! *erco!!" 'er2e&'

/!#'& *e"%!r& *er&!!" *H *e"%!r& o"2e"'r!2# e",# 2er'! *e"%!r&

o"2e"'r!2# 2&2'r!' er#&' er&*!!" !2#+"/!

1 U"'& *H

P!$! *H $#2e$#!!" 3 e!$!!" /!#'& !2! (HC+) !2! (N!OH) $!"

"e'r!+ (!&!$e2') M!2&!" $!+! '!&"% re!2# /!"% 'e+! 'er+e# $!&+&

$##2# $e"%!" H7O7 /!"% e&$#!" $#&&r *H "/! e&$#!" $#!2&!"

e2'r! e"'!"% e&$#!" !!" &+!# 'ere"'& %e+e&"% Ge+e&"% #"#

e"e"$!!" !9! 2&2'r!' !2! *ero2#$! 'e+! 'er&r!# e"4!$# !#r $!"

%!2 o2#%e" ; %e+e&"% (&#)

2 4 6 8 10 12

0

0.01

0.01

0.02Laju Pembentukan O2

Laju

Pembentukan

O2

D!r# %r!-# $#!'!2 $!*!' $#!'!!" !9! E",# er!$! *!$! *H

!2#& /!#'& 5 $!" *!$! *H 11 e",# e"%!+!# $e"!'&r!2#

7 U"'& o"2e"'r!2# 2&2'r!'

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0 0.5 1 1.5 2 2.5

0

0.01

0.02

0.03

Laju Pembentukan O2

Laju

PembentukanO2 (cm3/s

atau mL/s

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