enzim laporan praktikum biokimia
TRANSCRIPT
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
T
B 31@ 057 05 B0,3960
B 31@ 075 05 B00<
B 03=
7 V =π r2
.t 7 BV
T
B 31@ 057 @ B3,14
60
B 31@ 075 @ B 005
B 31@
3 V =π r2
.t 3 BV
T
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
T
B 31@ 057 0< B0,47
60
B 31@ 075 0< B000=
B 0@
7 V =π r2
.t 7 BV
T
B 31@ 057 10 B0,78
60
B 31@ 075 10 B 001
B 0>
3 V =π r2
.t 3 BV
T
B 31@ 057 1< B1,25
60
B 175 B007
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@ V =π r2
.t 3 BV
T
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
T
B 31@ 057 7> B
2,91
60
B 31@ 075 7> B 003
B 0@
7 V =π r2
.t 7 BV
T
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
T
B 31@ 057 5 B3,92
60
B 175 B00<
@ V =
π r
2
.t @ B
V
T
B 31@ 057 55 B4,31
60
B 03= B 00
5 V =π r2
.t 5 BV
T
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
D!+! *e"e"'&!" o"2e"'r!2# 2&2'r!' %r!-# /!"% $#$!*!' 2e*er'#
$#!'!2 H!+ #"# $#2e!!" !re"! $!'! $!" e!&r!'!" /!"% &r!"% !#
3 U"'& o"2e"'r!2# E",#
0 1 2 3 4
0
0.05
0.1
0.15
Laju Pembentukan O2 (cm3/s atau mL/s)
Laju
Pembentukan
O2 (cm3/s
atau mL/s
6eer!*! !+ /!"% e"/e!!" %r!-# *!$! *e"e"'&!" o"2e"'r!2#
2&2'r!' &r!"% !# /!#'& &r!"% 'e+#'#"/! *er#'&"%!" &r!"% !# $!+!
e"%%&"!!" !+!' $++
- Ke2#*&+!"
D!r# !2#+ *r!'#& /!"% !# +!&!" $#$!*!'!" !2#+ 2:
E",# er&*!!" 6#o!'!+#2!'or E",# 4&%! er*er!" e*erce*!'
re!2# '!"*! #&' ere!2#
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F!c'or ? -!c'or /!"% e*e"%!r&# er4! e",# /!#'&: Ko"2e"'r!2#
2&2'r!' Ko"2e"'r!2# E",# *H ('#"%!' e!2!!" ) S&&A$!"/!
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