. 2004 Aug 24;382(Pt 2):751–757. doi: 10.1042/BJ20040697

Table 1. Steady-state kinetic parameters and pK_a values for the thiol group of GSH of wild-type and mutants of adGSTD3-3 for the CDNB conjugation reaction at pH 6.5 and 25 °C.

The enzyme activities were measured at varying concentrations of CDNB and GSH in 0.1 M phosphate buffer, pH 6.5. The pK_a was obtained by using 0.1 M sodium acetate buffers (from pH 5.0 to pH 5.5) and 0.1 M potassium phosphate buffer (from pH 6.0 to pH 8.5).The reaction was monitored at 340 nm, ε=9600 M⁻¹·cm⁻¹.

Enzyme	k_cat (s⁻¹)	K_m^GSH (mM)	K_m^CDNB (mM)	k_cat/K_m^GSH (s⁻¹/mM)	k_cat/K_m^CDNB (s⁻¹/mM)	pK_a
Wild-type	35.4	0.27±0.05	0.14±0.01	130.69	245.95	6.36±0.11
I52A	16.9	6.50±0.61	0.39±0.03	2.60	43.72	6.78±0.14
S65A	29.8	1.22±0.12	0.34±0.06	24.50	87.18	6.89±0.23
R66A	3.3	5.10±0.40	0.22±0.04	0.64	14.59	7.23±0.18
M101A	63.0	1.08±0.09	0.19±0.01	58.48	337.12	6.10±0.07

Table 1. Steady-state kinetic parameters and pKa values for the thiol group of GSH of wild-type and mutants of adGSTD3-3 for the CDNB conjugation reaction at pH 6.5 and 25 °C.

Table 1. Steady-state kinetic parameters and pK_a values for the thiol group of GSH of wild-type and mutants of adGSTD3-3 for the CDNB conjugation reaction at pH 6.5 and 25 °C.