Frans_M_Everaerts_Isotachophoresis_378342.pdf

ENZYMATIC CONVERSION OF PYRUVATE 359
I0 20 30 40 50 ca 70
- time
Fig.15.7. Zone lengths of the various reaction constituents as a function of the reaction time in an
enzymatic reaction in which LDH is involved.
TABLE 15.8
CHANGE IN COMPOSITION OF A REACTION MIXTURE CAUSED BY THE ENZYME LACTATE
DEHY DROGENASE
Reaction time Zone length (mm)
(min)
Pyr uvate Lactate NADH NAD
0
10
30
40
60
70
80
90
100
120
27.4
27.3
24.3
23.3
22.0
21.7
20.9
21.1
25.0
20.4
1.4
2.7
4.2
4.9
6.5
7.3
7.9
7.1
2.9
8.7
~-____
17.6
16.2
11.3
9.5
6.1
5.0
3.7
5 .O
14.1
2.1
that are always present in the chemicals.
For this series of experiments, a mass balance was made for all reaction constituents.
In order to determine the amounts of pyruvate and NADH in nanomoles that fit a zone
length of 1 mm, the values obtained at t = 0, the reaction mixture before the conversion,
were applied, as these amounts are known exactly at this time. For lactate and NAD, a
calibration graph was constructed, which is quicker* than using the calibration constant
*Moreover, accurate data are not available for the determination of the calibration constant.
-
2.3
4.5
5.8
8.3
9.3
10.2
9.0
2.5
11.1