:

' .

:
)
( )( .
CoA-" - .
)" ( , 300-
, 10nM- 0.5mM- .
, " :

] [ H3
.
' ) (
) ( - .
PH ,
) ( .
.
.
. ] [C13
.
 :
:

)OVA(mg
0.072 0.0025
0.140 0.0050
0.230 0.0075
0.222 0.0100
0.268 0.0125

) ' 3 (

0.3
0.25
)O.D (595nm

0.2
0.15
y = 22.435x
0.1 2
R = 0.9477
0.05
0
0 0.005 0.01 0.015
)OVA (mg

" - . ". A = l C :


% )(mg/ml )(mg OD
2.32E-03 0.052 1:10-
50.54 2.51 1.25E-02 0.281
4.50 0.22 8.91E-05 0.002 1:10-
1.34E-03 0.03
44.96 2.23 1.11E-03 0.025 1:10-
0.85 4.23E-03 0.095 total
0.281
: = 1.25 10 2 mg -
22.43

1.25 10 2 mg
3
= 2.51 -
5 10 ml

2.51
' " 100 = 50.54% -
)( 2.51 + 0.22 + 2.23
 :

)(l
nmol Ach*H J&R
nmol nmol
min mg min ml min ml CPM )(l )(l )(l
- - 0.00 1978 60 30 10 - Control
)3*106( - 120000.00 26683 10 30 10 - Total
0.00 1821 55 30 10 5 5-
2956.97 7407.25 8297.92 7103 45 30 10 15 15-
6516.58 15394 10 30 10 50 50-
8956.93 1996.20 1486.34 2284 55 30 10 5 5-
2127.50 3292 45 30 10 15 15-
2374.74 6867 10 30 10 50 50-
4598.92 10249.42 10652.09 4171 55 30 10 5 5-
9904.07 8095 45 30 10 15 15-
10192.11 22961 10 30 10 50 50-

: 15- -
300nmol (cpm cpmcontrol ) 10 300(7103 1978) 10 nmol
= Act = = 8297.92
(cpmtotal cpmcontrol ) 5 min v E ml (26683 1978) 5 0.015 min ml

5- cpm- control-
.

Act nmol
= S.A min ml = 7407.25 = 2956.97 nmol -
mg 2 . 51 min mg
C protein
ml

.

mgenzyme
)(%
mgprotein
)(%
17.9 0.099
54.2 0.298
27.8 0.153

' -
mg enzyme S . A nmol min mg p 2956.57 100
= = 100% = 0.099%
mg protein 3 10 nmol min mg E
6
3 10 6

yield = 0.099 + 0.298 + 0.153 = 0.550% :

0.099
= . ratio % 100% = 17.9% -
0.099 + 0.298 + 0.153
 :

) 28:54:18':':'( . 3-
70:20:10 ' 10% '
20%-' .70% :
,
" ,
A ) (G
.
:. - ,
. "
.. -
, . . -
- .

, .0.550%:

3 10 6 nmol - ) (total-
min mg
total ) 3- ( . -
total ,12%- -
.0.550%
total- ""
, ) (80%
3- .
, 3- "
. , ""
"" , .

,
"
.

http://www.lsbu.ac.uk/biology/enztech/breakage.html