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The second screen was performed using the optimal substrate composition identified in the first screen.
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Reaction mixtures were prepared using the optimal conditions for catalysis reported previously for these substrates (27).
Then, stretch using the optimal.
Using this optimal substrate, the limit of detection (LOD) of arsenate was determined to be approximately 5 μg·l-1.
The optimal substrate was then used for (i) testing recombinantly expressed POMGnT1 mutant enzymes for activity, (ii) determining the kinetic parameters of the active POMGnT1 variants, and (iii) evaluating the possibility of correlating genotype with the obtained biochemical data.
Brinkworth et al have made use of the available crystal structures, molecular modelling and sequence analyses of kinases and substrates to predict the optimal substrate peptides [7].
As the optimal substrate stearoyl-ACP for the SSI2 desaturase was not present as the major component in E. coli, this enzyme used palmitoyl-ACP as an alternative substrate to produce palmitoleate.
The optimal substrate was sucrose.
The optimal substrate composition should satisfy the metabolic target function, while requiring as little oxygen and substrates as possible.
Here, using the PKD substrate antibody that specifically recognize the phosphorylated LXRXXp(S/T) motif, we show that ABF3 is phosphorylated in vivo in response to ABA on at least one SnRK2 optimal phosphorylation site.
As described in the methods section, the data sets of substrates and inhibitors were separated into a training and a test set using the D-optimal onion design (DOOD).
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