2.4.2 Measurement with pure Tmm
The amount of NADPH consumed and TMAO formed is assayed using the
previously reported methods. The Michaelis-Menten equation is used for
calculating Km and Vmax forTmm . The values for Km and Vmaxare calculated as 0.13 µM and 1.631 µM, respectively, when considering
NADPH consumed in the reaction. The values for Km and
Vmax are calculated as 0.08 µM and 0.85 µM,
respectively, when the amount of TMAO formed in the reaction is
monitored. Whereas activity measured through the current reported method
is lower (figure 4). The Kmvalues indicate the substrate and enzyme interaction, and a lower value
of Km suggests a better binding of Tmm with its
native substrate, trimethylamine. Thus, our method is more accurate
compared to NADPH-dependent assay.