Figure 1.
Reaction schemes for the tested enzymes.
(A) Reactions in the glucose metabolism catalyzed by GAPDH and PGK. In the forward reaction for GAPDH, conversion of substrate GAP is monitored directly by changes in NADH concentration. In the forward reaction for PGK, conversion of substrate ADP is monitored in a linked assay with forward reaction of GAPDH as a source for 1,3 BPG. In the reverse reaction for PGK, conversion of 3-PGA is monitored by the changes in NADH concentration in a linked assay with 1,3 BPG as a substrate for GAPDH [22]. (B) Reaction catalyzed by ACP. Hydrolysis of acylphosphate leads to the formation of a carboxylate and an inorganic phosphate.
Table 1.
Kinetic parameters of PGK, GAPDH and ACP in the absence or presence of the crowding agent Ficoll.
Table 2.
Crowding agent effects on the kinetic parameter, Km, of different enzymes.