93
Equation 14 can be rearranged to,
V V
e yo
= V
l-M
- 1
[I]
Cl] + Kl
+ Vi^H
(15)
This can be rewritten as,
Ve vo
ve=h
- i =
- 1
[I]
[I] + K,
(16)
Equation (16) can be linearized by a double reciprocal to give,
K_
Ve Vc
VU=H
*£=
- 1
ke=h ,
[I]
iKE=H
KE=h/K=H,
*1
(17)
Figure 22. If we let the observed elution volume, Ve, for [I] = 0 be
defined as V and from equation (13) we obtain,
Vq = Ve=h + V0 (IS)
This substitution into equation (15) yields an equation which can be ex
pressed in terms of experimental elution volumes for the enzyme. This
equation obviates the need to determine V0 or any column constant which
does not use the enzyme in its determination.
Ve Vc, VL^
[I]
LI] + Er
(19)