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AGUNG SUPRIflNTO PENGARUH SUMBER KARBON TE R H A D A P K E C E P A TA N P E R TU M B U H A N d a n K A N D U N G A N S TE R O ID K U L T U R K A L U S D IO S C O R E A PENTAPHYLLA L

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AGUNG SUPRIflNTO

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DISETUJUI OLEH PEMBIMBING :

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S e g a l a 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 o h S W T . a t a s s e g a l a r a h m a t d a n k a r u n i a n y a , s e h i n g g a

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

-d u l P E N G A R U H S U M B E R K A R B O N T E R H A D A P K E C E P A T A N P E R T U M B U H A N

D A N K A N D U N G A N S T E R O I D K U L T U R K A L U S D I O S C O R E A P E N T A P H Y L L A

L. g u n a m e m e n u h i t u g a s a k h i r m e n c a p a i g e l a r s a r j a n a

farraa-si p a d a F a k u l t a s F a r m a s i U n i v e r s i t a s A i r l a n g g a .

P a d a k e s e m p a t a n i n i p e n u l i s s a m p a i k a n u c a p a n t e r i m a

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

1. B a p a k DR. G u n a w a n I n d r a y a n t o d a n I b u D r a . L e s t a r i

R a h a y u M S s e l a k u d o s e n p e m b i m b i n g , y a n g t e l a h m e m b e r i

-k a n b i m b i n g a n d a n s a r a n - s a r a n .

2. B a p a k - I b u d o s e n p e n g u j i y a n g t e l a h b e r k e n a n m e m e r i k s a

d a n m e m b e r i k a n s a r a n .

3. P a r a s t a f L a b o r a t o r i u m B i o t e k n o l o g i F a k u l t a s F a r m a s i

U n i v e r s i t a s A i r l a n g g a y a n g b a n y a k m e m b e r i k a n b a n t u a n

s e l a m a p e n e l i t i a n .

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

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S e m o g a A l l a h S W T . m e m b e r i k a n b a l a s a n a t a s s e m u a b a n

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

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

m a n f a a t b a g i p e n g e m b a n g a n i l m u p e n g e t a h u a n .

S u r a b a y a , J u l i 1 9 9 0

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1 . K A T A P E N G A N T A R ... i

2. D A F T A R I S I ... i i i 3. D A F T A R T A B E L ... v

4. D A F T A R G A M B A R ... v i 5. D A F T A R I S T I L A H D A N S I N G K A T A N ... v i i 6 . B A B I P E N D A H U L U A N ... 1

1 . 1 . L a t a r B e l a k a n g ... 1

1 . 2 . T u j u a n P e n e l i t i a n ... 4

7. B A B II T I N J A U A N P U S T A K A ... 5

2 . 1 . K u l t u r J a r i n g a n ... 5

2 . 2 . M e d i a K u l t u r J a r i n g a n ... 6

2 . 3 . K o n d i s i L i n g k u n g a n K u l t u r ... 9

2 . 4 . S u m b e r K a r b o n ... 11

2 . 5 . K u l t u r K a l u s d a n k a r a k t e r i s t i k n y a ... 12

2 . 6 . K u l t u r J a r i n g a n s e b a g a i S u m b e r M e t a b o l i t S e k u n d e r ... 13

2 . 7 . T i n j a u a n T e n t a n g D i o s c o r e a p e n t a p h v l l a L. 15 1. K l a s i f i k a s i ... 15

2. N a m a D a e r a h ... 15

3. K a n d u n g a n D i o s c o r e a s p . . ... 15

N o . I s i H a l a m a n

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(7)

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

p a k a i p e n e l i t i a n ... 18

2. K o m p o s i s i m e d i a M u r a s h i g e d a n S k o o g y a n g d i

g u n a k a n p a d a p e n e l i t i a n ... 24

3. P e m e r i k s a a n m a k r o s k o p i k k a l u s D i o s c o r e a p e n t a

-p h v l l a L ... 25

4. P e n g a r u h s u m b e r k a r b o n p a d a b e r b a g a i k o n s e n

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

D i o s c o r e a p e n t a p h v l l a L ... 26

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

s t e r o i d k u l t u r k a l u s D i o s c o r e a p e n t a p h v l l a L. 27

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

r o l r e l a t i p t e r h a d a p k o l e s t e r o l k u l t u r k a l u s

D i o s c o r e a p e n t a p h v l l a L . 34

N o . T e k s H a l a m a n

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D A F T A R G A M B A R

1. K u r v a p e r t u m b u h a n s e l ... 13

2. G a m b a r h a s i l k r o m a t o g r a f i l a p i s t i p i s k a n d u n g

-a n k -a l u s D i o s c o r e -a p e n t -a p h v l l -a L ... 28

3. K r o m a t o g r a m e k s t r a k k a l u s D i o s c o r e a p e n t a p h v

-1 -1 a L. d e n g a n s u m b e r k a r b o n g l u k o s a 5 % .... 29

4. K r o m a t o g r a m e k s t r a k k a l u s D i o s c o r e a p e n t a p h v

-1 -1 a L. d e n g a n s u m b e r k a r b o n g l u k o s a 5 % y a n g

d i k o - e l u s i d e n g a n s t i g m a s t e r o l ( 5 : 1 ) 30

5. K r o m a t o g r a m e k s t r a k k a l u s D i o s c o r e a p e n t a p h v

-1 -1 a L. d e n g a n s u m b e r k a r b o n g l u k o s a 5 % y a n g

d i k o - e l u s i d e n g a n s i t o s t e r o l d a n k a m p e s t e r o l

( 5 : 1 ) ... 31

6 . G r a f i k k u r v a l i n i e r ( m g / m l V s a r e a p u n c a k ) 32

7. G r a f i k i n d e k s p e r t u m b u h a n d a n k a d a r s i t o s t e

-r o l k a l u s D i o s c o -r e a p e n t a p h v l l a L. p e n g a -r u h

s u m b e r k a r b o n ... . ... 33

N o . T e k s H a l a m a n

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B. A B e n z y l a m i n o p u r i n e ; B A P

2 , 4 D 2 , 4 - d i c h l o r o p h e n o x a c e t i c a c i d

I P i n d e k s p e r t u m b u h a n

D A A i n d o l e - 3 - a c e t i c a c i d

M S M u r a s h i g e d a n S k o o g

N A A a l f a - n a p h t h a l e n e a c e t i c a c i d

K L T K r o m a t o g r a f i L a p i s a n T i p i s

H P L C H i g h P r e s s u r e L i q u i d C h r o m a t o g r a p h y

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P e n e l i t i a n b e r i k u t d i m a k s u d k a n u n t u k m e n g e t a h u i

p e n g a r u h s u m b e r k a r b o n p a d a b e r b a g a i k o n s e n t r a s i t e r ­

h a d a p p e r t u m b u h a n d a n k a n d u n g a n k u l t u r k a l u s D i o s c o ­

r e a p e n t a p h v l l a L. D a r i p e n e l i t i a n m e n g e n a i i n i s i a s i

k a l u s D i o s c o r e a p e n t a p h v l l a L. d i k e t a h u i k e m a m p u a n

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

k a l u s t e r j a d i r a t a - r a t a 7 s a m p a i 14 h a r i s e t e l a h p e

-n a -n a m a -n e k s p l a -n d e -n g a -n p e r t u m b u h a -n y a -n g l a m b a t . (6 ).

T e l a h d i l a k u k a n p u l a o p t i m a s i m e d i a p e r t u m b u h a n d e

n g a n c a r a m e n g u b a h u b a h k o n s e n t r a s i h o r m o n p e r t u m b u h

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

o p t i m a l , y a i t u m e d i a M u r a s h i g e d a n S k o o g ( M S ) y a n g

d i k o m b i n a s i d e n g a n h o r m o n p e r t u m b u h a n k i n e t i n 2 p p m +

2 , 4 D 2 p p m d a n k i n e t i n 1 p p m + 2 , 4 D 4 p p m . K u l t u r

k a l u s y a n g d i d a p a t k a n p a d a p e m e r i k s a a n d e n g a n k r o m a

-t o g r a f i l a p i s -t i p i s -t e r d e -t e k s i a d a n y a s -t e r o l . (7)

O p t i m a s i m e d i a p e r t u m b u h a n d a p a t p u l a d i l a k u k a n

d e n g a n m e n g u b a h - u b a h k o n s e n t r a s i s u m b e r k a r b o n . P a d a

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

t u r j a r i n g a n a d a l a h : s u k r o s a , g l u k o s a , m a n n o s a , g a

-l a k t o s a , d a n -l a i n - -l a i n , y a n g m a s i n g - m a s i n g a k a n m e m ­

b e r i k a n r e s p o n p e r t u m b u h a n b e r b e d a t e r g a n t u n g p a d a

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3

P e n e l i t i a n t e n t a n g p e n g a r u h s u m b e r k a r b o n u n t u k

k e b u t u h a n n u t r i s i k u l t u r j a r i n g a n a g a r m e m b e r i k a n

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

b e b e r a p a j e n i s t a n a m a n , y a i t u p e n e l i t i a n R o k e m e t . a l .

d e n g a n m e n g u b a h - u b a h k o n s e n t r a s i s u k r o s a b e r h a s i l

m e n d a p a t k a n k a d a r d i o s g e n i n d a r i k u l t u r D i o s c o r e a

d e l t o i d e a 3 , 8 % b o b o t k e r i n g . K o n s e n t r a s i s u k r o s a y a n g

d i g u n a k a n a d a l a h 1 , 5 % (8 ).

P e n g a r u h k o n s e n t r a s i s u k r o s a y a n g d i g u n a k a n p a d a

p e r t u m b u h a n k u l t u r k a l u s u n t u k p r o d u k s i m e t a b o l i t s e

-k u n d e r , j u g a d i s e l i d i k i o l e h B e r l i n e t . a l . ( 1 9 8 6 ) ,

y a n g d e n g a n m e n a i k k a n k o n s e n t r a s i s u k r o s a d a r i 2%

m e n j a d i 1 0% m e n y e b a b k a n a k u m u l a s i b e t a l a i n p a d a

k u l t u r C h e n o p o d i u m r u b r u m s a m p a i 10 k a l i n y a . (9)

H a r i y a n t o ( 1 9 8 7 ) m e n e l i t i p e n g a r u h s u m b e r k a r b o n

t e r h a d a p p e r t u m b u h a n k u l t u r j a r i n g a n S o l a n u m m a m m o s u m

L. d a n S o l a n u m t u b e r o s u m L . , y a n g m e n u n j u k k a n h a s i l

t e r b a i k b e r t u r u t - t u r u t a d a l a h s u k r o s a , g l u k o s a d a n

l a k t o s a . (2 2 )

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

n u n j u k k a n b a h w a k o n s e n t r a s i s u m b e r k a r b o n m e m p e n g a

-r u h i k e c e p a t a n p e r t u m b u h a n p a d a k u l t u r j a r i n g a n t a

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1 . 2 . T u . iuan P e n e l i t i a n

P e n e l i t i a n b e r i k u t b e r t u j u a n u n t u k m e n g e t a h u i

p e n g a r u h s u m b e r k a r b o n g l u k o s a d a n s u k r o s a d e n g a n d e ­

n g a n m a s i n g - m a s i n g k o n s e n t r a s i 2%, 3 % d a n 5 % t e r h a d a p

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

k a l u s D i o s c o r e a p e n t a p h v l l a L. y a n g d i t a n a m p a d a m e

d i a M u r a s h i g e d a n S k o o g ( M S ) y a n g d i m o d i f i k a s i d e n g a n

p e n a m b a h a n h o r m o n p e r t u m b u h a n k i n e t i n 2 p p m d a n 2 , 4 D

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B A B

XX

T I N J A U A N P U S T A K A

2 . 1 . K u l t u r J a r i n g a n

K u l t u r j a r i n g a n d a p a t d i d e f i n i s i k a n s e b a g a i b a

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

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

p a d a s u a t u m e d i a b u a t a n d a n s e l - s e l n y a d a p a t t u m b u h

d a n m e n g a d a k a n p e m b e l a h a n . K u l t u r d a p a t b e r u p a k u l ­

t u r o r g a n t e r t e n t u y a n g t e l a h t e r d i f e r e n s i a s i a t a u

m a s s a s e l y a n g t a k t e r d i f e r e n s i a s i y a n g d i s e b u t k u l ­

t u r k a l u s . (2 )

M e t o d e k u l t u r j a r i n g a n t a n a m a n d i d a s a r i o l e h

t e o r i s e l d a r i S c h w a n ( 1 8 3 9 ) d a n S c h l e i d e n , y a n g m e

-n y a t a k a -n b a h w a s e l h e w a n d a n s e l t u m b u h a n m e r u p a k a n

s a t u a n b i o l o g i t e r k e c i l y a n g m a m p u m e l a k u k a n a k t i v i

-t a s h i d u p s e p e r -t i m e -t a b o l i s m e , r e p r o d u k s i d a n -t u m b u h .

M o r g a n m e n g e m u k a k a n b a h w a s e l j u g a m e m p u n y a i s i f a t

t o t i p o t e n s i , y a i t u s e m u a s e l t u m b u h a n m e m b a w a i n f o r

-m a s i g e n e t i k y a n g s a m a , d a n s e l d a l a m m e d i a d e n g a n

l i n g k u n g a n y a n g c o c o k p a d a h a k e k a t n y a m a m p u m e n g a d a ­

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

i n i y a n g m e n j a d i d a s a r b e r k e m b a n g n y a t e k n i k k u l t u r

j a r i n g a n . (1 0 )

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M e t o d e k u l t u r j a r i n g a n d a p a t d i m a n f a a t k a n u n t u k

t u j u a n - t u j u a n t e r t e n t u d a l a m b i d a n g f a r m a s i , a n t a r a

l a i n (2 ) :

1. P r o d u k s i s e n y a w a t e r t e n t u y a n g b e r g u n a d a l a m b i ­

d a n g f a r m a s i a t a u d a l a m b i d a n g m e d i k .

2. B i o t r a n s f o r m a s i s e n y a w a t e r t e n t u m e n j a d i s e n y a w a

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

d i k .

3. P r o d u k s i s e n y a w a - s e n y a w a s p e s i f i k

4. S e l e k s i g a l u r - g a l u r t a n a m a n u n g g u l u n t u k d i p a k a i

p a d a b i d a n g p e r t a n i a n d a n h o r t i k u l t u r a .

5. M u l t i p l i k a s i t a n a m a n s e c a r a c e p a t d a n s e r a g a m .

P e n e r a p a n m e t o d e k u l t u r j a r i n g a n i n i d i a w a l i o l e h

W h i t e ( 1 9 3 4 ) y a n g b e r h a s i l m e m b u a t k u l t u r j a r i n g a n

d a r i a k a r t o m a t (S o l a n u m l v c o p e r s i c u m ) . (1)

2 . 2 . M e d i a K u l t u r J a r i n g a n .

K o m p o s i s i d a r i m e d i a k u l t u r m e m e g a n g p e r a n a n y a n g

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

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

b u h a n , p r o l i f e r a s i d a n b i o s i n t e s a , k a r e n a m e d i a t e r

-s e b u t a k a n -s a n g a t b e r p e n g a r u h t e r h a d a p p r o -s e -s p e r t u m ­

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

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2 . M a k r o e l e r a e n a n o r g a n i k .

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

-t u h k a n u n t u k p e r t u m b u h a n m e l i p u t i ' N, P, M g , S,

K, C a , C l . N i t r o g e n d i b u t u h k a n d a l a m j u m l a h b e

-s a r , d i b e r i k a n d a l a m b e n t u k i o n n i t r a t d a n a t a u

a m m o n i u m . M a g n e s i u m d a n s u l f u r d i b e r i k a n d a l a m

b e n t u k M g S0 4 7 H2O. K a l i u m d i b e r i k a n d a l a m b e n t u k

K C 1 , K N O3 , K H2P O4 , d a l a m j u m l a h b e s a r p u l a . P e

-. n a m b a h a n C a C l2 2H2 O d a n C a f N O j ^ 4H2O a t a u b e n t u k

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

S e d a n g k a n k l o r i d a d a l a m b e n t u k K c l a t a u C a C l2 *

F o s f o r d a l a m b e n t u k N a l ^ P O ^ H2O a t a u K H2 P O4 . ( 3 , 8 )

3. M i k r o e l e m e n a n o r g a n i k

D i b u t u h k a n o l e h s e l t a n a m a n d a l a m j u m l a h k e c i l , N u t r i s i m i k r o e l e m e n y a n g d i p e r l u k a n s e l t a n a m a n

t i n g g i a d a l a h C u , Z n , M n , F e , B o d a n M o , b e b e r a p a

m e d i a j u g a m e n g a n d u n g C o d a n I ( 10 ) .

4. V i t a m i n

V i t a m i n d i p e r l u k a n d a l a m j u m l a h k e c i l , m e m p u n y a i

f u n g s i m e n g k a t a l i s a s i s t e m e n s i m . T h i a m i n m e r u p a

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

k u l t u r j a r i n g a n t a n a m a n . A s a m n i k o t i n a t d a n p i r i

(18)

-9

t a m b a h k a n p a d a b e b e r e p a m e d i a d e n g a n k o n s e n t r a s i

1 0 0 m g / l t , d i m a k s u d k a n s e b a g a i f a k t o r p e r t u m b u h a n .

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

a m i n o b e n z o a t , a s a m a s k o r b a t , b i o t i n , k h o l i n , s i a

n o k o b a l a m i n , a s a m f o l a t d a n r i b o f l a v i n (1 0 ).

5. Z a t P e n g a t u r T u m b u h

Z a t p e n g a t u r t u m b u h y a n g d i g u n a k a n u n t u k k e b a n y a k

-k a n -k u l t u r j a r i n g a n t a n a m a n t e r u t a m a -k u l t u r k a l u s

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

-r a n a n a n t a -r a l a i n t e -r h a d a p p e n g e m b a n g a n s e l , p e m

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

m e m p u n y a i p e r a n a n d a l a m p r o s e s p e m b e l a h a n s e l .

T e r m a s u k dalara k e l o m p o k a u k s i n y a n g s e r i n g d i g u n a ­

k a n a d a a h I A A , 2 , 4 D, N A A , s e d a n g k a n d a r i s i t o k i ­

n i n a d a l a h k i n e t i n , B A d a n z e a t i n (3).

2 . 3 . K o n d i s i L i n g k u n g a n k u l t u r

1 . p H .

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

b e r k i s a r a n t a r a p H 5 , 0 - 6 , 5 . M e n u r u t W h i t e p H

o p t i m u m p e r t u m b u h a n k u l t u r j a r i n g a n a d a l a h 5 , 4 ,

s e d a n g k a n m e n u r u t T u l l e c k e e t . a l . d a r i h a s i l

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

(19)

p e r t u m b u h a n a n t a r a 5 , 5 - 5 , 6 . B e b e r a p a m e d i a

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

w a k t u p e r t u m b u h a n p H d a p a t b e r u b a h . N a m u n p e n a m

-b a h a n n y a s a n g a t t e r -b a t a s , k a r e n a p a d a k o n s e n t r a s i

t i n g g i a k a n m e n g h a m b a t p e r t u m b u h a n k u l t u r ( 1 8 ) .

2. T e m p e r a t u r

M e n u r u t D e C a p i t e , t e m p e r a t u r o p t i m u m b e b e r a p a

k u l t u r j a r i n g a n l e b i h t i n g g i d i b a n d i n g k a n d e n g a n

t a n a m a n a s a l n y a , t e t a p i m e n u r u t R i k e r d a n H i l d e

-b r a n d t s a m a d e n g a n t a n a m a n a s a l n y a . S e c a r a u m u m

k u l t u r j a r i n g a n t a n a m a n a k a n t u m b u h d e n g a n b a i k

p a d a t e m p e r a t u r 20* - 28* C d e n g a n t e m p e r a t u r o p ­

t i m u m 26* - 27* C. T e t a p i b e b e r a p a k u l t u r j a r i n ­

g a n t i d a k m e n g a l a m i k e r u s a k a n p a d a t e m p e r a t u r 5*

-1 0 “ C. ( -1 8 )

3. P e n c a h a y a a n .

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

c a h a y a a n r e d u p , m e s k i p u n T o r r e y d a n R e i n e r t m e l a

-p o r k a n b a h w a k e c e -p a t a n t u m b u h d a l a m k e a d a a n g e l a p

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

B l a k e r l y d a n S t e w a r d m e l a p o r k a n p e n c a h a y a a n p a d a

k u l t u r j a r i n g a n H a p l o p a p p u s g r a v i l i s m e n g h a s i l k a n

(20)
(21)

s U R

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

d i o s g e n i n y a n g b e r b e d a (8 ).

K u l t u r k a l u s d a n k a r a k t e r i s t i k n y a

K u l t u r j a r i n g a n t a n a m a n y a n g p a l i n g u m u m a d a l a h

k u l t u r k a l u s , d i m a n a m e r u p a k a n s u a t u m a s a y a n g t i d a k

t e r d e f e r e n s i a s i d a r i s e l t a n a m a n . P e n e l i t i a n y a n g

d i l a k u k a n o l e h W h i t e d a n k e m u d i a n G a u t h e r e t d a n N o

-b e c o u r t p a d a t a h u n 1 9 3 0 m e m -b u k t i k a n -b a h w a k a l u s d a p a t

t e r b e n t u k d a r i j a r i n g a n t a n a m a n y a n g t e r i s o l a s i . K a

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

y a n g u m u m a d a l a h d e n g a n m e n a n a m b a g i a n t a n a m a n y a n g

t e l a h d i p o t o n g - p o t o n g d e n g a n u k u r a n t e r t e n t u , d i s t e

-r i l k a n d a n d i t a n a m p a d a m e d i a k u l t u -r y a n g s e s u a i ( 1 4 )

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

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

b u l a n . P a d a a k h i r n y a a k a n d i c a p a i u k u r a n y a n g o p t i ­

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

-s e b a b k a n m a k i n b e r k u r a n g n y a n u t r i s i a t a u d e h i d r a s i

m e d i a , t e r b e n t u k n y a m e t a b o l i t b e r a c u n , h a b i a n y a o k

-s i g e n -s e l i n t i j a r i n g a n ( 15 ) .

K u r v a p e r t u m b u h a n k a l u s s e l a l u m e n g h a s i l k a n b e n ­

t u k " S " ( s i g m o i d ) . K e c e p a t a n p e r t u m b u h a n m e n c a p a i

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13

f a s e s t a s i o n e r , k a r e n a s e l m e n g a l a m i m a t u r a s i d a n

p e r t u m b u h a n a k h i r n y a r a e n u r u n ( g a m b a r 1) ( 3 , 8 )

Jum

l*K Ml

G a m b a r 1 : K u r v a p e r t u m b u h a n s e l t a n a m a n

2 . 6 . K u l t u r J a r i n g a n s e b a g a i S u m b e r M e t a b o l i t S e k u n d e r

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

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

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

(23)
(24)
(25)

M E T O D O L O G I P E N E L I T I A N

3 . 1 . B a h a n .

3 . 1 . 1 . K a l u s

K a l u s D i o s c o r e a p e n t a p h v l l a L. d i p e r o l e h d a r i

l a b o t a r i u m B i o k t e k n o l o g i j u r u s a n B i o l o g i F a r

-raasi F a k u l t a s F a r m a s i U n i v e r s i t a s A i r l a n g g a .

3 . 1 * 2 . B a h a n K i m i a .

B a h a n kiraia y a n g d i g u n a k a n a d a l a h p r o d u k s i E.

M e r c k D a r m s t a d t d e n g a n d e r a j a t " p r o a n a l i s a " ,

k e c u a l i d i s e b u t k a n l a i n . A g a r y a n g d i g u n a k a n

a d a l a h B a c t o a g a r , D i f c o C e n t r i f r i e d , D i f c o

L a b o r a t o r i e s , D e t r o i t M i c h i g a n U S A . S u m b e r

k a r b o n y a n g d i g u n a k a n s u k r o s a d a n g l u k o s a

m o n o h y d r a t d a r i f e r a x l a b o r a t G M B H B e r l i n

(W e s t ).

3 . 1 . 3 . M e d i a p e r t u m b u h a n .

Y a n g d i g u n a k a n d a l a m p e n e l i t i a n i n i a d a l a h m e ­

d i a s t a n d a r M u r a s h i g e d a n S k o o g y a n g s e l a n j u t

-n y a d i s e b u t m e d i a s t a -n d a r M S ( t a b e l 2) d e n g a n

s u m b e r k a r b o n p a d a b e r b a g a i k o n s e n t r a s i ( t a b e l

1 ) d a n d e n g a n p e n a m b a h a n z a t p e n g a t u r t u m b u h

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17

k i n e t i n 2 p p m d a n 2 , 4 D 2 p p m .

3 . 2 . A l a t .

A l a t y a n g d i g u n a k a n d a l a m p e n e l i t i a n :

- L a m i n a r a i r f l o w c a b i n e t , u n t u k p e n g e r j a a n a s e p t i s

- A u t o c l a f , u n t u k s t e r i l i s a s i m e d i a

- p H m e t e r F i s h e r , u n t u k m e n g u k u r p H m e d i a

- M o n o p a n b a l a n c e

- O v e n

- C e n t r i f u g e

- A l a t - a l a t g e l a s ( e r l e n m e y e r , b e k e r g l a s s d l l . )

- A l a t - a l a t u n t u k p e n g e r j a a n s t e r i l ( p i n s e t , s p a t e l

d a n l a i n - l a i n ).

3 . 3 . C a r a K e r i a .

3 . 3 . 1 . P e m b u a t a n M e d i a

P e m b u a t a n m e d i a d a l a m p e n e l i t i a n i n i d i l a k u k a n

s e s u a i d e n g a n m e t o d a M u r a s h i g e d a n S k o o g d a l a m

B h o j w a n i 1 9 8 3 ( 3 ) . S e m u a b a h a n k i m i a k o m p o n e n

m e d i a M S d i b u a t d a n d i s i m p a n d a l a m l a r u t a n

s t o k . U n t u k m e m b u a t m e d i a d e n g a n v o l u m e 1 l i ­

t e r d i g u n a k a n c a r a s e b a g a i b e r i k u t :

M a s i n g - m a s i n g l a r u t a n s t o k d i p i p e t 10 m l d i

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-k o s a s e b a g a i s u m b e r -k a r b o n d e n g a n k o n s e n t r a s i

y a n g t e r t e n t u ( T a b e l 1) k e m u d i a n d i t a m b a h

a q u a d e s t s a m p a i v o l u m e m e n d e k a t i 1 0 0 0 m l s a m

b i l d i a d u k d a n p H d i a t u r p a d a 5 , 7 d e n g a n p e ­

n a m b a h a n N a O H 0 , 1 N a t a u H C 1 0 , 1 N. S e t e l a h

d i t a m b a h a g a r 1% m e d i a d i t a m b a h a q u a d e s t s a m ­

p a i v o l u m e t e p a t 1 0 0 0 m l . L a r u t a n l a l u d i p a

-n a s k a -n s a m b i l d i a d u k s a m p a i m e -n d i d i h d a -n j e r

-n i h . D a l a m k e a d a a n p a n a s d i t u a n g k a n k e d a l a m

b o t o l k u l t u r , k e m u d i a n d i t u t u p d e n g a n a l u m u

-n i u m f o i l d a -n d i i k a t d e -n g a -n t a l i . S e l a n j u t n y a

d i s t e r i l k a n s e c a r a a u t o c l a f p a d a s u h u 1 2 1 * C

s e l a m a 2 0 m e n i t .

T a b e l 1.

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

1

K o d e M e d i a S u m b e r K a r b o n K o n s e n t r a s i

S2 S u k r o s a 2%

S3 S u k r o s a 3 %

S5 S u k r o s a 5 %

G 2 G l u k o s a 2 %

G 3 G l u k o s a 3 %

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3 . 3 . 2 . P e n a n a m a n d a n P e m i n d a h a n K a l u s .

P e n a n a m a n d a n p e m i n d a h a n k a l u s d i l a k u k a n s e c a

r a a s e p t i k d a l a m k o t a k p e m i n d a h b e r a l i r a n u d a

-r a ( l a m i n a r a i r f l o w c a b i n e t ) y a n g t e l a h d i

-b e r s i h k a n d e n g a n a l k o h o l 7 0 % . K e m u d i a n l a m p u

U V d i n y a l a k a n s e l a m a 20 m e n i t , b o t o l k u l t u r

d i m a s u k k a n k e d a l a m l a m i n a r a i r f l o w c a b i n e t .

K e m u d i a n t u t u p b o t o l d i b u k a m u l u t b o t o l d i

-f l a m b i r . D e n g a n m e n g g u n a k a n s c a p e l s t e r i l k a ­

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

1 * 2 g r a m . M u l u t b o t o l s e t e l a h d i f l a m b i r di

t u t u p d e n g a n a l u m u n i u m f o i l . K a l u s d i p a n e n s e ­

t e l a h 4 m i n g g u .

3 . 3 . 3 . P e n g a m a t a n P e r t u m b u h a n K a l u s .

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

i n d e k s p e r t u m b u h a n s e b a g a i b e r i k u t :

b e r a t b a s a h k a l u s a k h i r

I p =

-b e r a t -b a s a h k a l u s a w a l

B e r a t b a s a h k a l u s a w a l m e r u p a k a n s e l i s i h

a n t a r a b e r a t m e d i a + k a l u s d e n g a n b e r a t

m e d i a . B e r a t k a l u s a k h i r d i t e n t u k a n

s e t e l a h k a l u s d i p a n e n .

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3 * 3 . 4 . E k s t r a k s i

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

a g a r y a n g m e n e m p e l . S e t e l a h d i k e r i n g k a n d a l a m

l e m a r i p e n g e r i n g p a d a s u h u 4 0 - 6 0 * C , d i s e r b u k

d a n d i h o m o g e n k a n . K e m u d i a n s e r b u k k a l u s d i

-t i m b a n g 1 g r a m , d i t a m b a h k l o r o f o r m 1 0 m l s e

-l a n j u t n y a d i v o r t e x d u a k a l i s e l a m a 5 m e n i t .

F i l t r a t d i s i s i h k a n d a n r e s i d u d i v o r t e x k e m b a

-l i d e n g a n 1 0 m -l k -l o r o f o r m . F i l t r a t d i k u m p u l

-k a n d a n d i u a p -k a n . H a s i l p e n g u a p a n f i l t r a t d i

-i d e n t -i f -i k a s -i d e n g a n K L T d a n IIPLC. R e s i d u d i

h i d r o l i s a d e n g a n 5 m l H C 1 2 N p a d a s u h u 1 0 0 ' C ,

s e l a m a d u a j a m . S e t e l a h d i n g i n d i n e t r a l k a n

d e n g a n N a O H 1 0 N. S e l a n j u t n y a d i v o r t e x d e n g a n

10 m l k l o r o f o r m s e l a m a d u a m e n i t . F a s e k l o r o ­

f o r m y a n g d i d a p a t d i u a p k a n . S e l a n j u t n y a

d i a n a l i s a s e c a r a k u a l i t a t i p d a n k u a n t i t a t i p

d e n g a n K L T d a n H P L C .

3 . 3 . 5 * A n a l i s a K u a l i t a t i p d e n g a n K L T .

A n a l i s a k u a l i t a t i p d e n g a n K L T d i l a k u k a n t e r h a ­

d a p e k s t r a k k l o r o f o r m d a r i k u l t u r k a l u s .

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(31)

D e t e k t o r : U V - V I S S p e c t r o f o t o m e t e r

S P G - 6 A V , p a d a 2 0 5 nm.

R e k o r d e r C h r o m a t o p a c C - R 3 A

K o n t r o l S y s t e m c o n t r o l l e r S C L -6A

I n j e k t o r I t

L o o p " d a r i M i l t o n C o m ­

p a n y R a n o , N e v a d a .

2. A n a l i s a s e c a r a k u a n t i t a t i p

2 . 1 . P e m b u a t a n k u r v a l i n i e r

Di t i m b a n g s e c a r a a n a l i t i k s t a n d a r k o

-l e s t e r o -l , d i b u a t p a d a b e r b a g a i k o n s e n ­

t r a s i d a l a m p e l a r u t m e t a n o l . P e n i m b a n g

-a n u n t u k m -a s i n g - m -a s i n g k o n s e n t r -a s i d i ­

l a k u k a n d u a k a l i . K o n s e n t r a s i y a n g

d i b u a t a d a l a h ( m g / m l ) ; 0 , 0 1 : 0 , 0 2 :

0 , 0 5 : 0 , 1 : 0 , 2 : Q j 4 : 0 , 8 . K o n s e n ­

t r a s i y a n g d i b u a t k e m u d i a n d i i n j e k s i k a n

p a d a H P L C , a k a n d i d a p a t d a t a l u a s p u n

-c a k . D a r i d a t a k o n s e n t r a s i d a n l u a s

p u n c a k d i l a k u k a n p e r h i t u n g a n a n a l i s a

k o r e l a s i d a n r e g r e s i . A k a n d i d a p a t p e r

-s a m a a n g a r i s l u r u s . D a r i p e r s a m a a n

g a r i s l u r u s t e r s e b u t , d i b u a t gra\fik

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(33)
(34)
(35)

4 . 2 . H a s i l P e r h i t u n g a n

T a b e l 4. P e n g a r u h S u m b e r K a r b o n p a d a b e r b a g a i k o n s e n t r a s i

t e r h a d a p i n d e k s p e r t u m b u h a n ( I P ) r a t a - r a t a k u l t u r

k a l u s D i o s c o r e a p e n t a p h y l l a L.

K o d e M e d i a I n d e k s p e r t u m b u h a n r a t a - r a t a p a d a m i n g g u k e 4 (x ± S D )

S2 3 , 0 2 + 1 , 4 4

S3 1 , 2 9 + 0 , 0 5

S5 1 ,76 + 0 , 2 9

G 2 1 , 8 2 + 0 , 3 2

G 3 1 ,69 + 0 , 4 6

(36)

27

4 . 3 . H a s i l P e m e r i k s a a n d e n g a n K L T .

T a b e l 5. P e n g a r u h S u m b e r k a r b o n t e r h a d a p k a n d u n g a n

s t e r o i d k u l t u r k a l u s D i o s c o r e a p e n t a p h v l l a L.

]

FASE CERAK

1

PEHBAHDING

1

G 2 G3 G5 S2 S3 S5

1

VARNA

PBNAHPAK

NODA

n-hexana : etil asetat

kolesterol

ungu

A n isaldehid

{ 80 : 20 j

sulf&t

squalen

kuning

A nisaldehid

sulfat

klorofori : letanol

diosgenin

kuning

Anisaldehid

(37)

A B C

K e t e r a n g a n :

A : E k s t r a k k a l u s D i o s c o r e a p e n t a -p h v l l a L .

B : P e m b a n d i n g k o l e s t e r o l

C : P e m b a n d i n g s q u a l e n

F a s e g e r a k : n - h e x a n a : e t i l a s e t a t ( 8 0 : 2 0 )

P e n a m p a k n o d a : A n i s a l d e h i d s u l f a t

W a r n a : u n g u

k D

______

K e t e r a n g a n :

A : E k s t r a k k a l u s D i o s c o r e a p e n t a -p h v l l a L .

D : P e m b a n d i n g d i o s g e n i n

F a s e g e r a k : K l o r o f o r m : m e t a n o l ( 9 5 : 5)

P e n a m p a k n o d a : A n i s a l d e h i d s u l f a t

W a r n a : k u n i n g

G a m b a r 2

(38)

4 . 4 . H a s i l p e m e r i k s a a n d e n g a n H P L C

1. A n a l i s a k u a l i t a t i f

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' 2 3 . 3 9 2 5 . 1 4 . 8 4 6 8

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C H K 0 M A T 0 P A C c ~ f t 3 A T I L E £

s a m p l e: M O 0 M E T H O D 6 1

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T O T A L 1 1 5 0 6 0 7 1 0 0

(40)
(41)

2 . 2 . H a s i l P e m b u a t a n k u r v a l i n i e r s t a n d a r k o l e s t e r o l

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a

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G a m b a r 6 . G r a f i k k u r v a l i n i e r ( m g / m l V s A r e a p u n c a k )

(42)

M U I K

PE RiJU&i a KA A H "U N IV E R S IT A S A IR L A N G G A "

S U R A B A Y A

33

K a d a r S t e r o l k a l u s D . p e n t a p h y l l a

P e n g a r u h s u m b e r k a r b o n

4

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5

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IP, L - ) Sitoi'iorol

r ; j.

G a m b a r 7 : G r a f i k I n d e k s p e r t u m b u h a n ( I P ) d a n k a d a r

s i t o s t e r o l r e l a t i p t e r h a d a p k o l e s t e r o l

( m g / m l ) k u l t u r k a l u s D i o c o r e a p e n t a p h y l l a

(43)

T a b e l 6 : P e n g a r u h s u m b e r k a r b o n p a d a b e r b a g a i k o n s e n t r a s i

t e r h a d a p k a d a r s i t o s t e r o l r e l a t i p t e r h a d a p k o

-l e s t e r o -l k u -l t u r k a -l u s D i o s c o r e a p e n t a p h v -l -l a L.

k o d e m e d i a

k a d a r s i t o s t e r o l r a t a - r a t a r e l a t i p t e r h a d a p k o l e s t e r o l d a l a m m g / g k a l u s k e r i n g

j u m l a h s a m p e l

(n)

S2 0 , 2 8 9 n = 2

S3 0, 1 6 4 n = 1

S 5 0 , 3 0 8 n = 2

G 2 0 , 4 3 3 n = 1

G 2 0 , 1 6 2 n = 2

(44)

B A

.

B

V

P E M B A H A S A N

P a d a p e n e l i t i a n i n i d i s e l i d i k i p e n g a r u h p e n g g u n a a n

s u m b e r k a r b o n s u k r o s a d a n g l u k o s a p a d a b e r b a g a i k o n s e n t r a ­

s i t e r h a d a p p e r t u m b u h a n d a n k a n d u n g a n k u l t u r k a l u s D i o s c o ­

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

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

P a d a k u l t u r k a l u s D i o s c o r e a p e n t a p h v l l a L. t e r j a d i

p e n c o k l a t a n ( b r o w n i n g ) d a n w a k t u t e r j a d i n y a t i d a k s a m a .

K a l u s D i o s c o r e a p e n t a p h v l l a L. p a d a u m u m n y a b e r w a r n a c o k

-l a t s a m p a i c o k -l a t k u n i n g a g a k k e p u t i h a n . J i k a b a g i a n k a ­

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

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

w a k t u t e r j a d i n y a p e n c o k l a t a n l e b i h l a m b a t . T i m b u l n y a p e n ­

c o k l a t a n d i s e b a b k a n o l e h t e r b e n t u k n y a s e n y a w a - s e n y a w a f e

-n o l i k y a -n g k e m u d i a -n m e -n u m p u k d i d a l a m m e d i a , s e h i -n g g a m e ­

d i a b e r w a r n a c o k l a t . K e a d a a n i n i a k a n m e n g h a m b a t p e r t u m ­

b u h a n . U n t u k m e n d e t e k s i k a n d u n g a n k a l u s D i o s c o r e a p e n t a ­

p h v l l a L. d i l a k u k a n a n a l i s a s e c a r a k u a l i t a t i p d a n k u a n t i

-t a -t i p , d e n g a n K L T d a n H P L C . P a d a a n a l i s a k u a l i t a t i p

d e n g a n K L T d i t e m u k a n a d a n y a s t e r o l d a l a m k a l u s D i o s c o r e a

p e n t a p h y l l a L. U n t u k m e m a s t i k a n m a c a m d a n j u m l a h s t e r o l

y a n g t e r d a p a t d a l a m k a l u s D i o s c o r e a p e n t a p h v l l a L. d i l a k u

(45)

k a n a n a l i s a s e c a r a k u a l i t a t i p d a n k u a n t i t a t i p d e n g a n H P L C .

P a d a a n a l i s a k u a n t i t a t i p u n t u k m e m a s t i k a n a t a u m e r a p e r t e g a s

s u a t u s e n y a w a d a p a t d i l a k u k a n d e n g a n m e n g g u n a k a n s t a n d a r

i n t e r n a l a t a u d e n g a n c a r a k o - e l u s i . D a r i a n a l i s a k u a n t i ­

t a t i p d a p a t d i t u n j u k k a n k a n d u n g a n k a l u s D i o s c o r e a p e n t a ­

p h v l l a L. a d a l a h s i t o s t e r o l , s t i g m a s e t r o l d a n a t a u k a m

-p e s t e r o l .

D a r i g r a f i k i n d e k s p e r t u m b u h a n r a t a - r a t a d a n k a d a r

s i t o s t e r o l k a l u s D i o s c o r e a p e n t a p h v l l a L. P e n g a r u h s u m b e r

k a r b o n p a d a b e r b a g a i k o n s e n t r a s i d i k e t a h u i b a h w a r e l a t i p

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

-n a m p a d a m e d i a d e n g a n s u m b e r k a r b o n g l u k o s a 5%, s e d a n g

k a d a r s i t o s t e r o l t e r t i n g g i r e l a t i p t e r j a d i p a d a k a l u s y a n g

d i t a n a m p a d a m e d i a d e n g a n s u m b e r k a r b o n g l u k o s a 2%. R o k e m

e t . a l . ( 1 9 8 5 ) dalain p e n e l i t i a n n y a m e l a p o r k a n b a h w a s e m a k i n

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

k u l t u r k a l u s D i o s c o r e a p e n t a p h v l l a L. s e m a k i n b e s a r p u l a

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

k o n s e n t r a s i s u m b e r k a r b o n t e r h a d a p k a d a r d i o s g e n i n t i d a k

m e n u n j u k k a n a p a k a h d e n g a n k e n a i k k a n k o n s e n t r a s i s u k r o s a

a k a n m e n a i k k a n k a d a r d a r i d i o s g e n i n . P a d a p e n e l i t i a n t e r

-s e b u t k a d a r d i o s g e n i n y a n g d i h a s i l k a n 3 , 8 % b e r a t k e r i n g

d e n g a n k o n s e n t r a s i s u k r o s a 1 , 5 % , s e d a n g p a d a k o n s e n t r a s i

(46)
(47)

z i m - e n z i m t e r s e b u t d i e k s p r e s i k a n t e t a p i t i d a k a d a p r e k u r

-s o r , t i d a k a d a n y a k e m a m p u a n p e n y i m p a n a n p r o d u k , a t a u t e r

j a d i n y a d e g r a d a s i p r o d u k y a n g m e n y e b a b k a n p r o d u k t e r t e n t u

(48)

B A B

VX

K E S I M P U L A N

D a r i h a s i l p e n e l i t i a n i n i d a p a t d i s i m p u l k a n :

1. M e d i a d e n g a n s u m b e r k a r b o n g l u k o s a p a d a k o n s e n t r a s i 5 %

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

k a l u s D i o s c o r e a p e n t a p h v l l a L. y a n g d i t a n a m p a d a m e d i a

M S y a n g d i k o m b i n a s i k a n d e n g a n h o r m o n k i n e t i n 2 p p m d a n

2 , 4 D 2 p p m .

2. A n a l i s a d e n g a n K L T d a n H P L C t e r h a d a p k a n d u n g a n s t e r o i d

k u l t u r k a l u s D i o s c o r e a p e n t a p h v l l a L. p e n g a r u h s u m b e r

k a r b o n g l u k o s a d a n s u k r o s a m e n u n j u k k a n a d a n y a s i t o s ­

t e r o l d a n s t i g m a s t e r o l d e n g a n k a d a r t e r t i n g g i p a d a

k o n s e n t r a s i g l u k o s a 2 % d a n 5 %

(49)

S A R A N - S A R A N

S e t e l a h d i l a k u k a n p e n e l i t i a n mengeriai p e n g a r u h s u m ­

b e r k a r b o n p a d a p e r t u m b u h a n d a n k a n d u n g a n k u l t u r k a l u s

D i o s c o r e a p e n t a p h y l l a L. p e r l u d i l a k u k a n p e n e l i t i a n l e b i h

l a n j u t m e n g e n a i p e n g a r u h k o m p o n e n m e d i a y a n g l a i n t e r h a d a p

p e r t u m b u h a n k a l u s D i o s c o r e a p e n t a p h y l l a L. m i s a l n y a s u m b e r

N i t r o g e n a t a u p u n s u m b e r f o s f a t .

(50)

R I N G K A S A N

D i l a k u k a n p e n e l i t i a n p e n g g u n a a n s u m b e r k a r b o n y a n g

b e r b e d a p a d a b e r b a g a i k o n s e n t r a s i u n t u k m e n d a p a t k a n m e d i a

p e r t u m b u h a n o p t i m a l k u l t u r k a l u s D i o s c o r e a p e n t a p h v l l a L.

p a d a m e d i a M u r a s h i g e d a n S k o o g ( M S ) y a n g d i k o m b i n a s i

d e n g a n h o r m o n p e r t u m b u h a n k i n e t i n 2 p p m d a n 2 , 4 D 2 p p m .

M e d i a d e n g a n s u m b e r k a r b o n g l u k o s a 5 % r e l a t i p m e m

-b e r i k a n i n d e k s p e r t u m -b u h a n t e r t i n g g i , s e d a n g m e d i a d e n g a n

s u m b e r k a r b o n g l u k o s a 2 % m e m b e r i k a n k a n d u n g a n s i t o s t e r o l

r e l a t i p t e r t i n g g i .

P e n c o k l a t a n t e r j a d i p a d a m e d i a p e r t u m b u h a n k u l t u r

k a l u s D i o s c o r e a p e n t a p h v l l a L. p a d a p e r c o b a a n m e d i a d e n g a n

s u m b e r k a r b o n s u k r o s a 3 % a d a y a n g m e m b e r i k a n p e r t u m b u h a n

t u n a s d a n d a u n ( t e r d i f e r e n s i a s i ) d a r i k u l t u r k a l u s D i o s c o ­

r e a p e n t a p h v l l a L .

A n a l i s a s e c a r a k u a l i t a t i p d a n k u a n t i t a t i p t e r h a d a p

k a n d u n g a n k a l u s D i o s c o r e a p e n t a p h v l l a L. t e r d e t e k s i a d a n y a

s i t o s t e r o l d a n s t i g m a s t e r o l , d e n g a n k a d a r t e r t i n g g i p a d a

m e d i a d e n g a n p e n a m b a h a n s u m b e r k a r b o n g l u k o s a 2 % d a n 5%.

(51)

1. I s n a e n i , 1 9 8 6 . O p t i m a s i P e m b e n t u k a n K a l u s d a n I d e n t i

-f i k a s i s t e r o i d d a r i s o l a n u m m a m m o s u m L, T e s i s , F a k u l ­

t a s P a s c a S a r j a n a , U n i v e r s i t a s A i r l a n g g a , S u r a b a y a , p.

1 - 1 0 1 2 - 3 7 .

2. I n d r a y a n t o , G, 1 9 8 6 , P r o s p e k K u l t u r J a r i n g a n T a n a m a n

p a d a B i d a n g F a r m a s i , B u l l e t i n I S F I 17 (1), p. 1 2 - 2 2 .

3. B h o j w a n i , S S a n d M K R a z d a n , 1 9 8 3 , P l a n t T i s s u e C u l t u r e

T h e o r y a n d P r a c t i s e , E l s e i v i e r , A m s t e r d a m - O x f o r d , N e w

Y o r k - T o k y o , P. 2 5 - 3 2 .

4. W e t h e r e l , D. F . , 1 9 7 6 , P e n g a n t a r P r o p a g a s i T a n a m a n

S e c a r a i n V i t r o , t e r j e m a h a n , P e n e r b i t I K I P S e m a r a n g

P r e s s p . 4 5.

5. I n d r a y a n t o , G, 1 9 8 9 , A p l i k a s i B i o t e k n o l o g i S e l T a n a m a n

U n t u k B i d a n g F a r m a s i , U n i v e r s i t a s A i r l a n g g a , S u r a b a y a ,

p. 1-1 0 .

6 . R e t n o S a r i , 1 9 8 7 , S k r i p s i D r a . F a r m a s i , F a k u l t a s F a r ­

m a s i U n i v e r s i t a s A i r l a n g g a , S u r a b a y a .

7. A n y K o o s b u d i w a t i , 1 9 9 0 , S k r i p s i D r a . F a r m a s i F a k u l t a s

F a r m a s i U n i v e r s i t a s A i r l a n g g a , S u r a b a y a .

8 . R o k e m J. S. et. a l ., 1 9 8 5 , M e t h o d a F o r I n c r e a s i n g

D i o s e g e n i n P r o d u c t i o n b y D i o s c o r e a C e l l i n S u s p e n s i o n

C u l t u r e , J o u r n a l o f N a t u r a l P r o d u c t s , 4 8 (2), p. 2 1 0

-222

.

(52)
(53)

17. H e y n e , K. 1 9 5 0 , D e N u t t i g e P l a n t e n v a n I n d o n e s i a I, 3e

D r u k , N . V . V i t g e v e r i j v a n H o e v e ’s G r a v e n h a g e , B a n d u n g ,

p. 4 5 4 - 4 6 1 .

1 8. P u h a n , Z. a n d M a r t i n S . M . , 1 9 7 5 , T h e I n d u s t r i a l P o t e n

-s i a l o f P l a n t C e l l C u l t u r e , N a t . R e -s . C o u n c . o f C a n a ­

d a . 1 1 5 9 5 p. 1 5 - 1 9 , 23, 24.

19. T j o n d r o n e g o r o , P. D. e t . a l . , 1 9 8 6 , U s a h a P e n i n g k a t a n

K a d a r D i o s g e n i n p a d a D i o s c o r e a s p p . d e n g a n C a r a K u l t u r

I n V i t r o , I n s t i t u t P e r t a n i a n B o g o r , p. 4 2 - 4 7 .

20. L e m b a g a B i o l o g i N a s i o n a l - L I P I , 1 9 7 7 , U b i - u b i a n , B o ­

g o r , p . 3 9 .

2 1. C l a u s , E . P . 1 9 6 1 , P h a r m a c o g n o s y , 4 t h E d i t i o n , L e a

F e b i g e r : P h i l a d e l p h i a , p. 1 2 9 - 1 3 1 , 1 3 4 .

2 2. H a r i y a n t o , W, 1 9 8 7 , S k r i p s i , D r s . F a r m a s i , F a k u l t a s

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