; , 808 ڿ صȿ

 

(1) 1 ; ڿ

 

 

 

   <>

        赿 ; mouse 301g 1

         ; abs. EtOH 0.1ml in 1% CMC 0.3ml

         ; abs. EtOH 0.1ml 30 , ÷ 3

                 ( ; 20mg in 0.4ml CMC Ź)

        ÷ 30 , ä

 

   < >

        (ڿø ϰ ÷๰ ) ; 0.23%

         ; 0.16%(30.4% )

        808 ; 0.19%(17.4% )

         ; 0.30% (30.4% )

 

   <>

     ȿ ϳ mouse 1 ϿǷ Ȯ.

     ׷ ⼺ .

 

(2) 2 ; ڿúȿ(̿, ADH) Ȱȿ

 

   <>

        complex solution (++)(2.5ml)

   . acetaldehyde(2mM);0.011mg/2.5ml

       (0.4ml in 25ml buffer ߿ 2.5ml )

   . KCI(0.1M);18.65mg/2.5ml (0.93mg in 2.5ml buffer)

   . Tris-HCI buffer 0.1M(pH 8.0) 25ml

complex solution -NAD+(0.5mM) 100, sample()100, ()100 ALDHȿ Ǵ ADHȿ 100unit 100 쿩 ϰ 25ɿ 1а , 340nm O.D.

 

(3) 3 : 󵵺 ȿ Ȱ ȿ

ȿ 󵵺 , ADHȰ ġ 0.5mg/ml 󵵿 73.4% Ͽ 1mg/ml 󵵿 86.3% Ͽ.(Fig.2) ALDHȰ ġ 0.5mg/ml 󵵿 4.6% Ͽ.(Fig. 3)

 

0.5mg/ml 󵵿 ź ƼƮ˵ ȭŰ ALDHȰ ƼƮ˵ () ƼƮ ȭŰ ALDHȰ ϰ Ű ȴ.

ALDH

activity

(Abs at 340nm)

1.2

1.0

0.8

0.5

0.4

0.2

0

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

         0            0.5           1

Concentration(mg/ml)

 

ADH

activity

(Abs at 340nm)

1.0

0.8

0.5

0.4

0.2

0

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

         0            0.5           1

 

Concentration(mg/ml)

 

Fig 2. 󵵺 ADHȿ             Fig. 3 󵵺 ALDHȿ

 

 

 

<>

808 1mg/ml Ͽ ADH ȿ , 808 7.2 ȿ Ÿ.(Fig. 1)

 

ADH

activity

(Abs at 340nm)

0.8

0.5

0.4

0.2

0

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

               808

 

Concentration(mg/ml)

 

 

                Fig. 1  808 ADH Ȱ ȿ

 

⹰ 󵵿 Alcohol Dehydrogenase Activity

 

ź : 16M NAD : 20mM  ȿ : 10unit

 

(340nm )

 

Con

0.50%

0.75%

1.00%

1.50%

1ȸ

0.0920.102

0.5750.588

0.7840.788

1.0271.032

1.4431.448

2ȸ

0.0910.100

0.5740.584

0.7940.804

0.9951.014

1.4641.471

3ȸ

0.0940.103

0.5650.572

0.7910.800

0.9950.800

1.4651.473

 

ȭ

 

Con

0.50%

0.75%

1.00%

1.50%

1ȸ

0.010

0.013

0.004

0.005

0.005

2ȸ

0.009

0.010

0.010

0.019

0.007

3ȸ

0.009

0.007

0.009

0.013

0.008

 

The effect of extraction on Yeast acohol dehydrogenase

 

 

Reaction rate

Relative activity

Control

0.910-2

100%

0.50%

1.210-2

133.30%

0.75%

1.010-2

111.10%

1.00%

1.610-2

177.80%

1.50%

0.710-2

77.80%

 

 

ȿȰ

 

ȿ Ȱ ⹰

C2H50H+NADCH3CHO+NADH+H ̿

, ڿ ȭ Ͽ 340nm NADH

ADH ȿȰ ġ Ͽ.

 

ͱ

 

Τ٣ ; ٥, ٥808 - Ը

 

(1) С1 ; 顢 뫳-

 

   <۰>

        ڪ ; mouse 301g 1

         ; abs. EtOH 0.1ml in 1% CMC 0.3ml

         ; abs. EtOH 0.1ml 30 , 3 Ϣ

                 (Ϣ ; 20mg in 0.4ml CMC )

        30 ,

 

   < ̿>

        (- 横 ڪ 横ʪ) ; 0.23%

        ٥ ; 0.16%(30.4% )

        ٥808 ; 0.19%(17.4% )

         ; 0.30% (30.4% ʥ)

 

   <̿>

     ٥ mouse 1 ŪêΪ

     Ǫ⡢ 몯骤

 

(2) 2 ; -(ALDH, ADH) ˭

 

   <۰>

        complex solution (a+b+c) (2.5ml)

   a. acetaldehyde(2mM);0.011mg/2.5ml

       (0.4ml in 25ml buffer ªê 顢 2.5ml )

   b. KCI(0.1M);18.65mg/2.5ml (0.93mg in 2.5ml buffer)

   c. Tris-HCI buffer 0.1M(pH 8.0) 25ml

߾ complex solution -NAD+(0.5mM) 100, sample()100, ׶()100 ALDHȪȪ ADH100unit ׶ 100˪Ȫ ʥ 25ɪ 1 , 340nm O.D.

 

(3) 3 : ٥ ܬ ˭

٥ ܬ ۪̿, ADHª 0.5mg/ml ˪ ٪ 73.4% ʥơ 1mg/ml ˪ 86.3% ʥ(Fig.2)

ALDHª 0.5mg/ml ˪ 4.6% ʥ(Fig. 3)

 

ê 0.5mg/ml ǫ뫢뫳-򫢫ȫǫҫɪ ߫ ALDH ȫǫҫ(ڪ)򫢫ë߫ ߫ ALDH ˭ʥ몳Ȫ ҡ

٥ALDH

activity

(Abs at 340nm)

1.2

1.0

0.8

0.5

0.4

0.2

0

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

         0            0.5           1

Concentration(mg/ml)

 

٥ADH

activity

(Abs at 340nm)

1.0

0.8

0.5

0.4

0.2

0

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

         0            0.5           1

 

Concentration(mg/ml)

 

Fig 2. ٥ ܬ ADHʥ             Fig. 3 ٥ ܬ ALDHʥ

 

 

 

<̿>

٥ ٥808 1mg/ml Ūê ADH ʥ 򪷪 , ٥᪬ ٥808 7.2 ˭ ު覆(Fig. 1)

 

ADH

activity

(Abs at 340nm)

0.8

0.5

0.4

0.2

0

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

       ٥       ٥808

 

Concentration(mg/ml)

 

 

                Fig. 1  ٥ ٥808 ADH ʥ

٥ ڪ ˪ Alcohol Dehydrogenase Activity

 

뫢뫳- : 16M NAD : 20mM  : 10unit

 

̿(340nmǪ )

 

Con

0.50%

0.75%

1.00%

1.50%

1

0.0920.102

0.5750.588

0.7840.788

1.0271.032

1.4431.448

2

0.0910.100

0.5740.584

0.7940.804

0.9951.014

1.4641.471

3

0.0940.103

0.5650.572

0.7910.800

0.9950.800

1.4651.473

 

ܨ

 

Con

0.50%

0.75%

1.00%

1.50%

1

0.010

0.013

0.004

0.005

0.005

2

0.009

0.010

0.010

0.019

0.007

3

0.009

0.007

0.009

0.013

0.008

 

The effect of extraction on Yeast acohol dehydrogenase

 

 

Reaction rate

Relative activity

Control

0.910-2

100%

0.50%ڪ

1.210-2

133.30%

0.75%ڪ

1.010-2

111.10%

1.00%ڪ

1.610-2

177.80%

1.50%ڪ

0.710-2

77.80%

 

 

 

Ȫ ڪ Ǫ

C2H50H+NADCH3CHO+NADH+H

, - ߫ ת 340nm NADH ʥҪ

ڪ ADH ª Ъ