Fig. 8: Biochemical properties of recombinant TreS-Mak of Estrella lausannensis.

a The activity of trehalose synthase domain (TreS) of bifunctional TreS-Mak protein was first conducted at 30 °C pH 8 with 200 mM trehalose in presence of 0, 1, and 10 mM of manganese chloride (MnCl₂). The interconversion of trehalose into maltose and subsequent release of glucose (red) were inferred by using amyloglucosidase assay method. The TreS activity is expressed as μmol of maltose/min/mg of protein. b The effect of nucleoside triphosphate on TreS activity was determined by measuring the interconversion of trehalose into maltose in the presence of increasing concentration of ATP (0–20 mM) and 200 mM trehalose. c The apparent K_m value (n = 3) for trehalose was determined in the absence of ATP and 1 mM of MnCl₂ by measuring the interconversion of increase concentration of trehalose (0–200 mM) into maltose. d Maltokinase activity domain was inferred by measuring the release of ADP during phosphorylation of maltose (20 mM) into M1P in presence of 0, 1, 3, and 10 mM of MnCl₂. The Mak activity is expressed as μmol of ADP released/min/mg of protein. e The effects of divalent cation Mn²⁺, Ni²⁺, Co²⁺, Zn²⁺, Cu²⁺, Ca²⁺, and Mg²⁺ at 10 mM and f ATP, CTP, GTP, and UTP nucleotides on Mak activity were determined and expressed as relative percentage of maximum activity. Data are presented as individual data points with error bar denoting standard deviation of n ≥ 3 independent experiments.