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In both cases, the substrate adopted a non-native conformation, and PscF and YscF substrates were disordered at the N-terminus.
Two members of the superfamily have been reported to catalyse C C bond formation [ 16], whereby in both cases the substrate bears a coenzyme A ester moiety.
In both cases the substrate conversion time has a minimum at a specific ratio of the concentrations of the individual enzymes.
From this perspective in both cases, the substrate is embedded through the cofactor on one side and coordinated through the Zn2+ ion.
However, in both cases, the substrate oxidation produces carboxylic acids (d-gluconic acid and 3,3-carboxy cis, cis-muconic acid, respectively, Figure 1 a, reactions 1 and 2).
Similar(55)
In both cases, the width of the substrate was taken as 4000 nm.
There are interesting parallels to be drawn with other systems, most notably in the PrnB enzyme mentioned above and in indole alkaloid biosynthesis, 47) because in both cases the involvement of the substrate amine group in the reaction mechanism is likewise implicated.
In both cases, the SuperSignal West Pico chemoluminescent substrate from Pierce was used in line with the manufacturer's recommendations.
In both cases the luminescence increased with increasing substrate concentration in an exponential manner until a plateau was reached at 40 µM coelenterazine for Gluc and 1570 µM luciferin for FFluc, with 40 µM and 470 µm chosen as working concentrations.
In all cases, the substrate assimilation processes are controlled by Rayleigh-distillation-type kinetics, and the isotopic composition of the product is not constant but intrinsically variable.
In all cases, the substrate consumption was <20%, the linearity being ensured within this time interval even at the lowest substrate concentration.
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