While the study provides significant structural insights, there are some gaps in the exploration of functional dynamics. For example, the kinetic assays focus on steady-state parameters (e.g., k_cat/K_m) but do not address how conformational changes in the ATEYS loop dynamically regulate substrate access. Could the authors discuss whether time-resolved methods like stopped-flow spectroscopy or molecular dynamics simulations were considered to link structural flexibility with enzymatic kinetics? Additionally, the study demonstrates enhanced reactivity of the E493A mutant toward non-native substrates, yet it stops short of exploring its structural basis. Were cryo-EM or crystallographic data obtained for this mutant to confirm whether the altered activity corresponds to conformational changes?
