Studies have shown that endophytes are capable of metabolizing in vitro most substrates found in plants, and produce enzymes including proteases, amylases, phenol oxidases, lipases, laccases, polyphenol oxidases, cellulases, mannanases, xylanases, and pectin lyase [ 53, 54].
In general, the V806E exchange resulted in an increase in Km for most substrates (except retinaldehyde) and in a decrease in kcat.
In addition, most substrates showed that contact angle hystereses were in the range of 0° ~ 3° with pure cleaned mercury which means that the mercury can be easily roll off from the surface.
Despite the fact that most substrate molecules in a reaction were uncarboxylated, almost all carboxylated FIXQ/S molecules were carboxylated many times.
Few compounds such as the dimethyl- (8) and diisopropylethoxysilane (9) reacted readily at 30°C whereas most substrates necessitated elevated temperatures (65°C 90°C) in order to react with the F−.
Based on rapid direct enzyme-catalyzed reduction of the mediators and clear color changes we selected DCPIP, MG, and Thi, which show distinct absorption changes between their oxidized and reduced forms in the visible range, as the most promising substrates in these assays for further characterization.
As suggested by the reviewers, we have now measured the effects of PKA on its most relevant substrates in Hh signaling – Gli2 and Gli3, and have added these new data to the revised manuscript. 1) PKA is known to inhibit Gli2 translocation to the cilia tip, a process required for Gli2 activation (Tuson et al., 2011).
Most substrate-binding assays used in studying transporters, such as surface plasma resonance (SPR), isothermal titration calorimetry (ITC), or scintillation proximity assay (SPA), provide an apparent dissociation constant, K d,app, which is neither K d,in nor K d,out, but is a weighted average value of both (Zhang et al. 2015).
For each enzyme, activity was calculated relative to the assay in which the most substrate conversion was observed (i.e. the latter assay being set to 100%%).
