Identifikasi Enzim Lipoksigenease dari Beberapa Varietas Kacang Tanah (Arachis hypogaea)

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lDENTlFlKASl ENZIM EIPOKSIGENASE DARl BEBERAPW

VARlETAS KACANG TANAW

(Arachis

hypogaea)

Oleh

ASWATI ELIANA

1 9 8 9

FAKULTAS TEKWOLOOI PERTANIAN INSTITUT PERTANIAN BOGOR

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FAKULTAS TEKNOLOGI PERTANIAN INSTITUT PERTANIAN BOGOR

IDENTIFIKASI ENZIM LIPOKSIGENASE DARI BEBERAPA

VARIETAS KACANG TANAH

C A r ~ c h i s hypogQea>

O l e h

ASWATI _ELIANA

F 21.1282

D i l a h i r k a n d i M a g e l a n g , t a n g g a l 2 Desember 1965

t a n g g a l l u l u s : !O _~ _~ _:t(Jg$_~ _~ _b _i

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A s w a t i E l i a i ~ ~ a , F 2 1 . 1 2 8 2 . l d e n t i f i k a s i Enzim L i p n k s i g e n a s e d a r i B e b e r a p a V a r i e t a s K a c a n g 'l'anah ( A r a c h i s hypogaea). D i

bawah b i m b i i ~ g a n U r . T r . M . Aman W i r a k a r t a k u s u m a h , MSc.,

I r . B . A . S u s i l o S a n t o s a , MS. d a n D r . I r . Djoko S . D a m a r d j a t i ,

MSc.

RI NGKASAN

O k s i d a s i l e m a k m e r u p a k a n m a s a l a h s e r i u s y a n g d a p a t m e m - p e n g a r u h i mutu k a c a n g t a n a h d a n p r o d u k o l a h a n n y a . Enzim l i p o k s i g e n a s e m e r u p a k a n s a l a h s a t u k a t a l i s p e n t i n g y a n g d a - p a t m e n g k a t a l i s a o k s i d a s i asam l e m a k t i d a k j e n u h membentuk s e n y a w a h i d r o p e r o k s i d a . H i d r o p e r o k s i d a i n i d a p a t t e r u r a i m e n j a d i a s a m , k e t o n d a n a l d k h i d r a n t a i p e n d e k y a n g b e r p e r a n d a l a m p e m b e n t u k a n f l a v o r y a n g t i d a k d i s u k a i . S e l a i n i t u h i d r o p e r o k s i d a j u g a d a p a t b e r i n t e r a k s i d e n g a n p r o t e i n , asam

a m i n o d a n p e p t i d a m e n g h a s i l k a n s e n y a w a v o l a t i l d e n g a n b a u t i d a k e n a k y a n g s a n g a t m e n y e n g a t d a n d a p a t menurunkan n i l a i g i z i p r o d u k l e m a k k a c a n g t a n a h .

Dalam p e n e l i t i a n i n i d i a m a t i b e b e r a p a s i f a t d a s a r e n z i m l i p o k s i g e n a s e k a c a n g t a n a h . Dengan d i k e t a h u i n y a s i f a t - s i f a t d a n k a r a k t e r i s t i k l i p o k s i g e n a s e d a p a t d i u s a h a k a n p e n c e g a h a n a k t i v i t a s e n z i m i n i selama p r o s e s p e n g o l a h a n .

F r a k s i p r o t e i n t e r b e s a r d a l a m k a c a n g t a n a h a d a l a h f r a k -

s i g l o b u l i n , 4 1 . 3 - 4 8 . 1 p e r s e n d a r i t o t a l p r o t e i n y a n g t e r - e k s t r a k . S e d a n g k a n f r a k s i p r o t e i n d e n g a n a k t i v i t a s l i p o k s i - g e n a s e t i n g g i a d a l a h f r a k s i a l b u m i n , 4 0 . 1 - 5 3 . 8 p e r s e n t o t a l a k t i v i t a s . B e r d a s a r k a n h a s i l i n i l a h i s o l a s i e n z i m l i p o k s i - g e n a s e d i l a k u k a n t e r h a d a p f r a k s i a l b u m i n .

P u r i f i k a s i e n z i m l i p o k s i g e n a s e d e n g a n ammonium s u l f a t 4 0 - 6 0 p e r s e n j e n u h m e n i n g k a t k a n a k t i v i t a s s p e s i f i k m e n j a d i

1 . 9 - 2 . 6 k a l i a k t i v i t a s s p e s i f i k e n z i m k a s a r . P e m i s a h a n l i p o k s i g e n a s e d e n g a n g e l f i l t r a s i S E p h a d e x G-150 m e n g h a s i l - kan t i g a p u n c a k a k t i v i t a s u n t u k m a s i n g - m a s i n g g a l u r d e n g a n a k t i v i t a s s p e s i f i k 6 . 1 - 9 2 . 4 k a l i a k t i v i t a s s p e s i f i k e n z i m k a s a r

.

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l u r , y a l t u 18(101) ( H f U . 2 8 ) u n t u k v a r l e t a s i;a,?ah d a n 76000 ( H f 0 2 9 ) u n t u k g a l u r 1509 dart 1 5 1 2 .

Enzim l i p o k s i g e n a s e k a c a n g t a n a h t e r d e n a t u r a s i s e t e l a h pemanasan 711 "C s e l a m a 30 m e n i t u n t u k v a r i e t a s Ga.iah d a n 1 5 m e n i t u n t u k g a l u r 1509 d a n 1 5 1 2 . E n e r g i a k t i v a s i l i p o k s i g e - n a s e d a r i v a r i e t a s G a j a h l e b i h r e n d a h d a r i p a d a l i p o k s i g e n a - s e d a r i g a l u r 1509 darr i 5 1 2 , y a i t u 1 9 0 8 3 k a l / M o l . S e d a n g k a n e n e r g i a k t i v a s i l i p o k s i g e n a s e d a r i g a l u r 1509 d a n 1512 h a m p i r s a m a . y a i t u 25446 d a n 24780 k a l / M o l . H a s i l i n i menunjukkan bahwa l i p o k s i g e n a s e k a c a n g t a n a h v a r i e t a s G a j a h r e l a t i f l e b i h s t a b i l t e r h a d a p p a n a s d a r i p a d a g a l u r 1509 d a n 1 5 1 2 , k a r e n a b i l a e n e r g i a k t i v a s i l e b i h r e n d a h b e r a r t i p e n g a r u h p e r u b a h a n s u h u t e r h a d a p n i l a i k j u g a l e b i h k e c i l . Enzim i n i mempunyai a k t i v i t a s t i n g g i p a d a pH 6 - 7 , d a n t e r d e n a t u r a s i p a d a pH d i bawah 3 _{a t a u d i a t a s 1 0 .}

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KATA PENGANTAR

P u j i d a n s y u k u r p e n u l i s p a n j a t k a n k e h a d i r a t A l l a h SWT., k a r e n a h a n y a d e n g a n Rahmat-Nya p e n u l i s d a p a t n e n y e l e s a i k a n p e n e l i t i a n d a n p e n u l i s a n s k r i p s i .

S k r i p s i i n i d i s u s u n b e r d a s a r k a n h a s i l p e n e l i t i a n d i B a l a i P e n e l i t i a n Tanaman P a n g a n , Sukamandi s e l a m a d e l a p a n b u l a n m u l a i b u l a n A g u s t u s 1 9 8 8 h i n g g a M a r e t 1989.

P a d a k e s e m p a t a n i n i p e r k e n a n k a n l a h p e n u l i s menyampikan u c a p a n t e r i m a k a s i h kepada :

1. Bapak i b u d a n k e l u a r g a t e r c i n t a , a t a s d o r o n g a n n y a s e l a m a

p e n u l i s b e l a j a r d i IPB.

2 . D r . I r . M . Aman W i r a k a r t a k u s u m a h , MSc., s e l a k u d o s e n

pembimbing u t a m a a t a s p e n g a r a h a n d a n bimbingan s e l a m a p e n e l i t i a n h i n g g a t e r s u s u n n y a s k r i p s i i n i

3 . _{D r .} _{I r . D j o k o S .} _{D a m a r d j a t i ,} _MSc., _{s e l a k u pembimbimg}

p e n d a m p i n g d a n k e t u a Kelompok P e n e l i t i K i m i a , BPTP S u k a m a n d i

,

y a n g t e l a h memberi k e s e m p a t a n p e n u l i s u n t u k m e l a k u k a n p e n e l i t i a n d i l a b . K i m i a , BPTP Sukamandi s e r t a

p e n g a r a h a n d a n b i m b i n g a n s e l a m a p e n e l i t i a n d a n p e n y u s u n a n s k r i p s i .

4 . I r . B . A . S u s i l o S a n t o s a , MS., s e l a k u pembimbing

p e n d a m p i n g y a n g t e l a h b a n y a k memberikan b i m b i n g a n d a n p e n g a r a h a n s e l a m a p e n e l i t i a n d a n p e n y u s u n a n s k r i p s i .

5 . Karyawan l a b . Kimia BPTP S u k a m a n d i , yang t e l a h m e n b a n t u

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6 . Teman-teman s e p e r j u a n g a n d i S u k a m a n d i , y a n g t e l a h memberi s e m a n g a t selama p e n e l i t i a n .

7 . Semua p i h a k y a n g t e l a h membantu p e n u l i s .

P e n u l i s m e n y a d a r i bahwa t u l i s a n i n i m a s i h b e l u m s e m p u r n a . S a r a n d a n k r i t i k pembaca s a n g a t p e n u l i s h a r a p k a n .

A k h i r k a t a s e m o g a h a s i l p e n e l i t i a n d a n t u l i s a n i n i d a p a t b e r m a n f a a t b a g i s e m u a p i h a k y a n g m e m e r l u k a n i n f o r m a s i i n i .

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DAFTAR IS1

KATA PENGANTAR

. . .

DAFTAR I S 1

. . .

DAFTAR TABEL

. . .

DAFTAR GAMBAR

. . .

DAFTAR LAMPIRAN

I

.

PENDAHULUAN

. . .

...

I1

.

TINJAUAN PUSTAKA

A

.

KACANG TANAH

...

B

.

LIPOKSIGENASE

C

.

PURIFIKASI LIPOKSIGENASE

...

D

.

AKTIVITAS LIPOKSIGENASE

...

E

.

STABILITAS ENZIM

...

F

.

KINETIKA ENZIM

...

I11

.

BAHAN DAN METODA

...

A

.

ALAT DAN BAHAN

...

B

.

NETODA PENELITIAN

. . .

1

.

P e r s i a p a n S a m p e l

2

.

F r a k s i n a s i P r o t e i n K a c a n g T a n a h

. . .

3

.

_{E k s t r a k s i d a n I s o l a s i E n z i m L i p o k s i g e n a s e}

.

4

.

A n a l i s a A k t i v i t a s L i p o k s i g e n a s e

...

5

.

P e n e n t u a n P r o t e i n

. . .

6

.

P e n g a r u h Suhu T e r h a d a p A k t i v i t a s Enzim

...

7 . S t a b i l i t a s E n z i m T e r h a d a p pH

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9 .

Penentuan

P o l a

Protein

. . .

10

.

Penentuan Berat Molekul

. . .

IV

.

HASIL DAN PEMBAHASAN

. . .

A

.

KADAR AIR. LEMAK DAN PROTEIN

. . .

B

.

AKTIVITAS LIPOKSIGENASE DARI BERBAGAI FRAKSI

PROTEIN

. . .

C

.

. . .

D

.

BERAT MOLEKUL LIPOKSIGENASE

. . .

E

.

STABILITAS ENZIM TERHADAP pH

...

F

.

STABILITAS ENZIM TERHADAP PANAS

G

.

KINETIKA ENZIM

...

V

.

KESIMPULAN DAN SARAN

...

VI

.

DAFTAR PUSTAKA

...

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DAFTAR TABEL

T a b e l 1 . K o m p o s i s i k i m i a b i j i k a c a n g t a n a h

. . .

5 T a b e l 2 . K o m p o s i s i a i r , lemak dan p r o t e i n k a c a n g [image:9.523.35.456.92.559.2]

t a n a h

. . .

3 5

...

T a b e l 3 . F r a k s i - f r a k s i p r o t e i n k a c a n g t a n a h 36

T a b e l 4 . A k t i v i t a s l i p o k s i g e n a s e d a r i b e r b a g a i

f r a k s i p r o t e i n

...

3 9 T a b e l 5 . N i l a i Rf f r a k s i p r o t e i n k a c a n g t a n a h

v a r i e t a s G a j a h

...

4 1

T a b e l 6 . N i l a i Rf f r a k s i p r o t e i n k a c a n g t a n a h

G a l u r 1 5 0 9

...

4 2

T a b e l 7 . N i l a i R f f r a k s i p r o t e i n k a c a n g t a n a h

G a l u r 1 5 1 2

...

4 3

T a b e l 8 . A k t i v i t a s l i p o k s i g e n a s e p a d a t i a p t a h a p

p u r i f i k a s i

...

4 5

T a b e l 9 . P o l a p r o t e i n e l u e n S e p h a d e x 6-150

...

5 5 T a b e l 1 0 . N i l a i Rf d a n b e r a t n o l e k u l l i p o k s i g e n a s e

k a c a n g t a n a h

...

5 7

Tabek 11. P e r s a m a a n g a r i s d a n k o r e l a s i l a j u k e r u s a k a n e n z i m l i p o k s i g e n a s e k a c a n g t a n a h p a d a b e r b a g a i

...

s u h u 6 1

T a b e l 1 2 . K o n s t a n t a k e c e p a t a n r e a k s i l i p o k s i g e n a s e

. . .

k a c a n g t a n a h 6 2

T a b e l 1 3 . Nilai Km d a n Vmaks l i p o k s i g e n a s e k a c a n g

...

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DAFTAR GAMBAR

[image:10.527.35.463.93.730.2]

. . .

Gambar 1 . Penampang m e l i n t a n g b i j i k a c a n g t a n a h Gambar 2 . K e c e p a t a n k e r u s a k a n enzim p a d a b e r b a g a i

s u h u

...

Gambar 3 . P e n g a r u h s u h u t e r h a d a p s t a b i l i t a s e n z i m

Gambar 4 . S t a b i l i t a s enzim t e r h a d a p pH

. . .

Gambar 5 . P e n g a r u h k o n s e n t r a s i s u b s t r a t t e r h a d a p

....

k e c e p a t a n r e a k s i y a n g d i k a t a l i s a e n z i m Gambar 6 . " L i n e w e a v e r - B u r k p l o t " d a r i l / V t e r h a d a p

1/[S] y a n g d i p e r g u n a k a n u n t u k e v a l u a s i

g r a f i k Km d a n Vmaks

. . .

Gambar 7 . Metoda f r a k s i n a s i p r o t e i n

...

Gambar 8 . Bagan a l i r p u r i f i k a s i enzim l i p o k s i g e n a s e

...

Gambar 9 . P o l a p r o t e i n d a r i f r a k s i p r o t e i n k a c a n g t a n a h v a r i e t a s G a j a h

...

Gambar 1 2 . P o l a p r o t e i n d a r i f r a k s i p r o t e i n k a c a n g

t a n a h G a l u r 1 5 0 9

...

Gambar 13. P o l a p r o t e i n d a r i f r a k s i p r o t e i n k a c a n g t a n a h g a l u r 1512

...

Gambar 1 4 . pH optimum l i p o k s i g e n a s e k a c a n g t a n a h

Gambar 1 5 . P e m i s a h a n l i p o k s i g e n a s e k a c a n g t a n a h

. . .

v a r i e t a s G a j a h d e n g a n S e p h a d e x 6-150

g a l u r 1 5 0 9 d e n g a n S e p h a d e x G-150

...

g a l u r 1512 d e n g a n S e p h a d e x 6-150

. . .

Gambar 18. P o l a p r o t e i n e l u e n g e l f i t r a s i S e p h a d e x G-150

. . .

Gambar 1 9 . Kurva s t a n d a r b e r a t m o l e k u l d e n g a n SDS-PAGE Gambar 2 0 . P e n g a r u h pH t e r h a d a p a k t i v i t a s l i p o k -

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lDENTlFlKASl ENZIM EIPOKSIGENASE DARl BEBERAPW

VARlETAS KACANG TANAW

(Arachis

hypogaea)

Oleh

ASWATI ELIANA

1 9 8 9

FAKULTAS TEKWOLOOI PERTANIAN INSTITUT PERTANIAN BOGOR

(105)

FAKULTAS TEKNOLOGI PERTANIAN INSTITUT PERTANIAN BOGOR

IDENTIFIKASI ENZIM LIPOKSIGENASE DARI BEBERAPA

VARIETAS KACANG TANAH

C A r ~ c h i s hypogQea>

O l e h

ASWATI _ELIANA

F 21.1282

D i l a h i r k a n d i M a g e l a n g , t a n g g a l 2 Desember 1965

t a n g g a l l u l u s : !O _~ _~ _:t(Jg$_~ _~ _b _i

(106)

A s w a t i E l i a i ~ ~ a , F 2 1 . 1 2 8 2 . l d e n t i f i k a s i Enzim L i p n k s i g e n a s e d a r i B e b e r a p a V a r i e t a s K a c a n g 'l'anah ( A r a c h i s hypogaea). D i

bawah b i m b i i ~ g a n U r . T r . M . Aman W i r a k a r t a k u s u m a h , MSc.,

I r . B . A . S u s i l o S a n t o s a , MS. d a n D r . I r . Djoko S . D a m a r d j a t i ,

MSc.

RI NGKASAN

O k s i d a s i l e m a k m e r u p a k a n m a s a l a h s e r i u s y a n g d a p a t m e m - p e n g a r u h i mutu k a c a n g t a n a h d a n p r o d u k o l a h a n n y a . Enzim l i p o k s i g e n a s e m e r u p a k a n s a l a h s a t u k a t a l i s p e n t i n g y a n g d a - p a t m e n g k a t a l i s a o k s i d a s i asam l e m a k t i d a k j e n u h membentuk s e n y a w a h i d r o p e r o k s i d a . H i d r o p e r o k s i d a i n i d a p a t t e r u r a i m e n j a d i a s a m , k e t o n d a n a l d k h i d r a n t a i p e n d e k y a n g b e r p e r a n d a l a m p e m b e n t u k a n f l a v o r y a n g t i d a k d i s u k a i . S e l a i n i t u h i d r o p e r o k s i d a j u g a d a p a t b e r i n t e r a k s i d e n g a n p r o t e i n , asam

a m i n o d a n p e p t i d a m e n g h a s i l k a n s e n y a w a v o l a t i l d e n g a n b a u t i d a k e n a k y a n g s a n g a t m e n y e n g a t d a n d a p a t menurunkan n i l a i g i z i p r o d u k l e m a k k a c a n g t a n a h .

Dalam p e n e l i t i a n i n i d i a m a t i b e b e r a p a s i f a t d a s a r e n z i m l i p o k s i g e n a s e k a c a n g t a n a h . Dengan d i k e t a h u i n y a s i f a t - s i f a t d a n k a r a k t e r i s t i k l i p o k s i g e n a s e d a p a t d i u s a h a k a n p e n c e g a h a n a k t i v i t a s e n z i m i n i selama p r o s e s p e n g o l a h a n .

F r a k s i p r o t e i n t e r b e s a r d a l a m k a c a n g t a n a h a d a l a h f r a k -

s i g l o b u l i n , 4 1 . 3 - 4 8 . 1 p e r s e n d a r i t o t a l p r o t e i n y a n g t e r - e k s t r a k . S e d a n g k a n f r a k s i p r o t e i n d e n g a n a k t i v i t a s l i p o k s i - g e n a s e t i n g g i a d a l a h f r a k s i a l b u m i n , 4 0 . 1 - 5 3 . 8 p e r s e n t o t a l a k t i v i t a s . B e r d a s a r k a n h a s i l i n i l a h i s o l a s i e n z i m l i p o k s i - g e n a s e d i l a k u k a n t e r h a d a p f r a k s i a l b u m i n .

P u r i f i k a s i e n z i m l i p o k s i g e n a s e d e n g a n ammonium s u l f a t 4 0 - 6 0 p e r s e n j e n u h m e n i n g k a t k a n a k t i v i t a s s p e s i f i k m e n j a d i

1 . 9 - 2 . 6 k a l i a k t i v i t a s s p e s i f i k e n z i m k a s a r . P e m i s a h a n l i p o k s i g e n a s e d e n g a n g e l f i l t r a s i S E p h a d e x G-150 m e n g h a s i l - kan t i g a p u n c a k a k t i v i t a s u n t u k m a s i n g - m a s i n g g a l u r d e n g a n a k t i v i t a s s p e s i f i k 6 . 1 - 9 2 . 4 k a l i a k t i v i t a s s p e s i f i k e n z i m k a s a r

.

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l u r , y a l t u 18(101) ( H f U . 2 8 ) u n t u k v a r l e t a s i;a,?ah d a n 76000 ( H f 0 2 9 ) u n t u k g a l u r 1509 dart 1 5 1 2 .

Enzim l i p o k s i g e n a s e k a c a n g t a n a h t e r d e n a t u r a s i s e t e l a h pemanasan 711 "C s e l a m a 30 m e n i t u n t u k v a r i e t a s Ga.iah d a n 1 5 m e n i t u n t u k g a l u r 1509 d a n 1 5 1 2 . E n e r g i a k t i v a s i l i p o k s i g e - n a s e d a r i v a r i e t a s G a j a h l e b i h r e n d a h d a r i p a d a l i p o k s i g e n a - s e d a r i g a l u r 1509 darr i 5 1 2 , y a i t u 1 9 0 8 3 k a l / M o l . S e d a n g k a n e n e r g i a k t i v a s i l i p o k s i g e n a s e d a r i g a l u r 1509 d a n 1512 h a m p i r s a m a . y a i t u 25446 d a n 24780 k a l / M o l . H a s i l i n i menunjukkan bahwa l i p o k s i g e n a s e k a c a n g t a n a h v a r i e t a s G a j a h r e l a t i f l e b i h s t a b i l t e r h a d a p p a n a s d a r i p a d a g a l u r 1509 d a n 1 5 1 2 , k a r e n a b i l a e n e r g i a k t i v a s i l e b i h r e n d a h b e r a r t i p e n g a r u h p e r u b a h a n s u h u t e r h a d a p n i l a i k j u g a l e b i h k e c i l . Enzim i n i mempunyai a k t i v i t a s t i n g g i p a d a pH 6 - 7 , d a n t e r d e n a t u r a s i p a d a pH d i bawah 3 _{a t a u d i a t a s 1 0 .}

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KATA PENGANTAR

P u j i d a n s y u k u r p e n u l i s p a n j a t k a n k e h a d i r a t A l l a h SWT., k a r e n a h a n y a d e n g a n Rahmat-Nya p e n u l i s d a p a t n e n y e l e s a i k a n p e n e l i t i a n d a n p e n u l i s a n s k r i p s i .

S k r i p s i i n i d i s u s u n b e r d a s a r k a n h a s i l p e n e l i t i a n d i B a l a i P e n e l i t i a n Tanaman P a n g a n , Sukamandi s e l a m a d e l a p a n b u l a n m u l a i b u l a n A g u s t u s 1 9 8 8 h i n g g a M a r e t 1989.

P a d a k e s e m p a t a n i n i p e r k e n a n k a n l a h p e n u l i s menyampikan u c a p a n t e r i m a k a s i h kepada :

1. Bapak i b u d a n k e l u a r g a t e r c i n t a , a t a s d o r o n g a n n y a s e l a m a

p e n u l i s b e l a j a r d i IPB.

2 . D r . I r . M . Aman W i r a k a r t a k u s u m a h , MSc., s e l a k u d o s e n

pembimbing u t a m a a t a s p e n g a r a h a n d a n bimbingan s e l a m a p e n e l i t i a n h i n g g a t e r s u s u n n y a s k r i p s i i n i

3 . _{D r .} _{I r . D j o k o S .} _{D a m a r d j a t i ,} _MSc., _{s e l a k u pembimbimg}

p e n d a m p i n g d a n k e t u a Kelompok P e n e l i t i K i m i a , BPTP S u k a m a n d i

,

y a n g t e l a h memberi k e s e m p a t a n p e n u l i s u n t u k m e l a k u k a n p e n e l i t i a n d i l a b . K i m i a , BPTP Sukamandi s e r t a

p e n g a r a h a n d a n b i m b i n g a n s e l a m a p e n e l i t i a n d a n p e n y u s u n a n s k r i p s i .

4 . I r . B . A . S u s i l o S a n t o s a , MS., s e l a k u pembimbing

p e n d a m p i n g y a n g t e l a h b a n y a k memberikan b i m b i n g a n d a n p e n g a r a h a n s e l a m a p e n e l i t i a n d a n p e n y u s u n a n s k r i p s i .

5 . Karyawan l a b . Kimia BPTP S u k a m a n d i , yang t e l a h m e n b a n t u

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6 . Teman-teman s e p e r j u a n g a n d i S u k a m a n d i , y a n g t e l a h memberi s e m a n g a t selama p e n e l i t i a n .

7 . Semua p i h a k y a n g t e l a h membantu p e n u l i s .

P e n u l i s m e n y a d a r i bahwa t u l i s a n i n i m a s i h b e l u m s e m p u r n a . S a r a n d a n k r i t i k pembaca s a n g a t p e n u l i s h a r a p k a n .

A k h i r k a t a s e m o g a h a s i l p e n e l i t i a n d a n t u l i s a n i n i d a p a t b e r m a n f a a t b a g i s e m u a p i h a k y a n g m e m e r l u k a n i n f o r m a s i i n i .

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DAFTAR IS1

KATA PENGANTAR

. . .

DAFTAR I S 1

. . .

DAFTAR TABEL

. . .

DAFTAR GAMBAR

. . .

DAFTAR LAMPIRAN

I

.

PENDAHULUAN

. . .

...

I1

.

TINJAUAN PUSTAKA

A

.

KACANG TANAH

...

B

.

LIPOKSIGENASE

C

.

...

D

.

AKTIVITAS LIPOKSIGENASE

...

E

.

STABILITAS ENZIM

...

F

.

KINETIKA ENZIM

...

I11

.

BAHAN DAN METODA

...

A

.

ALAT DAN BAHAN

...

B

.

NETODA PENELITIAN

. . .

1

.

P e r s i a p a n S a m p e l

2

.

F r a k s i n a s i P r o t e i n K a c a n g T a n a h

. . .

3

.

_{E k s t r a k s i d a n I s o l a s i E n z i m L i p o k s i g e n a s e}

.

4

.

A n a l i s a A k t i v i t a s L i p o k s i g e n a s e

...

5

.

P e n e n t u a n P r o t e i n

. . .

6

.

P e n g a r u h Suhu T e r h a d a p A k t i v i t a s Enzim

...

7 . S t a b i l i t a s E n z i m T e r h a d a p pH

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9 .

Penentuan

P o l a

Protein

. . .

10

.

Penentuan Berat Molekul

. . .

IV

.

HASIL DAN PEMBAHASAN

. . .

A

.

KADAR AIR. LEMAK DAN PROTEIN

. . .

B

.

AKTIVITAS LIPOKSIGENASE DARI BERBAGAI FRAKSI

PROTEIN

. . .

C

.

. . .

D

.

BERAT MOLEKUL LIPOKSIGENASE

. . .

E

.

STABILITAS ENZIM TERHADAP pH

...

F

.

STABILITAS ENZIM TERHADAP PANAS

G

.

KINETIKA ENZIM

...

V

.

KESIMPULAN DAN SARAN

...

VI

.

DAFTAR PUSTAKA

...

(112)

DAFTAR TABEL

T a b e l 1 . K o m p o s i s i k i m i a b i j i k a c a n g t a n a h

. . .

5 T a b e l 2 . K o m p o s i s i a i r , lemak dan p r o t e i n k a c a n g [image:112.523.35.456.92.559.2]

t a n a h

. . .

3 5

...

T a b e l 3 . F r a k s i - f r a k s i p r o t e i n k a c a n g t a n a h 36

T a b e l 4 . A k t i v i t a s l i p o k s i g e n a s e d a r i b e r b a g a i

f r a k s i p r o t e i n

...

3 9 T a b e l 5 . N i l a i Rf f r a k s i p r o t e i n k a c a n g t a n a h

v a r i e t a s G a j a h

...

4 1

T a b e l 6 . N i l a i Rf f r a k s i p r o t e i n k a c a n g t a n a h

G a l u r 1 5 0 9

...

4 2

T a b e l 7 . N i l a i R f f r a k s i p r o t e i n k a c a n g t a n a h

G a l u r 1 5 1 2

...

4 3

T a b e l 8 . A k t i v i t a s l i p o k s i g e n a s e p a d a t i a p t a h a p

p u r i f i k a s i

...

4 5

T a b e l 9 . P o l a p r o t e i n e l u e n S e p h a d e x 6-150

...

5 5 T a b e l 1 0 . N i l a i Rf d a n b e r a t n o l e k u l l i p o k s i g e n a s e

...

5 7

Tabek 11. P e r s a m a a n g a r i s d a n k o r e l a s i l a j u k e r u s a k a n e n z i m l i p o k s i g e n a s e k a c a n g t a n a h p a d a b e r b a g a i

...

s u h u 6 1

T a b e l 1 2 . K o n s t a n t a k e c e p a t a n r e a k s i l i p o k s i g e n a s e

. . .

k a c a n g t a n a h 6 2

T a b e l 1 3 . Nilai Km d a n Vmaks l i p o k s i g e n a s e k a c a n g

...

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DAFTAR GAMBAR

[image:113.527.35.463.93.730.2]

. . .

Gambar 1 . Penampang m e l i n t a n g b i j i k a c a n g t a n a h Gambar 2 . K e c e p a t a n k e r u s a k a n enzim p a d a b e r b a g a i

s u h u

...

Gambar 3 . P e n g a r u h s u h u t e r h a d a p s t a b i l i t a s e n z i m

Gambar 4 . S t a b i l i t a s enzim t e r h a d a p pH

. . .

Gambar 5 . P e n g a r u h k o n s e n t r a s i s u b s t r a t t e r h a d a p

....

k e c e p a t a n r e a k s i y a n g d i k a t a l i s a e n z i m Gambar 6 . " L i n e w e a v e r - B u r k p l o t " d a r i l / V t e r h a d a p

1/[S] y a n g d i p e r g u n a k a n u n t u k e v a l u a s i

g r a f i k Km d a n Vmaks

. . .

Gambar 7 . Metoda f r a k s i n a s i p r o t e i n

...

Gambar 8 . Bagan a l i r p u r i f i k a s i enzim l i p o k s i g e n a s e

...

Gambar 9 . P o l a p r o t e i n d a r i f r a k s i p r o t e i n k a c a n g t a n a h v a r i e t a s G a j a h

...

Gambar 1 2 . P o l a p r o t e i n d a r i f r a k s i p r o t e i n k a c a n g

t a n a h G a l u r 1 5 0 9

...

Gambar 13. P o l a p r o t e i n d a r i f r a k s i p r o t e i n k a c a n g t a n a h g a l u r 1512

...

Gambar 1 4 . pH optimum l i p o k s i g e n a s e k a c a n g t a n a h

. . .

v a r i e t a s G a j a h d e n g a n S e p h a d e x 6-150

g a l u r 1 5 0 9 d e n g a n S e p h a d e x G-150

...

g a l u r 1512 d e n g a n S e p h a d e x 6-150

. . .

Gambar 18. P o l a p r o t e i n e l u e n g e l f i t r a s i S e p h a d e x G-150

. . .

Gambar 1 9 . Kurva s t a n d a r b e r a t m o l e k u l d e n g a n SDS-PAGE Gambar 2 0 . P e n g a r u h pH t e r h a d a p a k t i v i t a s l i p o k -