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For supporting videos illustrating the ease of operation, see Supplementary Movies 1, 2. The corresponding organolithium reagent (1.2 eq) was added over a mixture of substrate (1.0 eq) and catalyst (1.5 3 mol%) at RT for 10 min.
In a typical catalytic run, 5 % H2O2 (18 g) was added dropwise to a suspension of the catalyst (Au 1 mol %) in a mixture of substrate (1 mmol) and deionized water (10 g) contained in a 50 mL two-neck round bottom flask, kept at 90 °C using a thermostatic oil bath.
Wild-type Arabidopsis thaliana, T-DNA mutants and transgenic plants (all of them in Col-0 accession) were grown either in pots containing a mixture of substrate and vermiculite (3∶1) or aseptically in Petri dishes containing Murashige and Skoog media supplemented with 1% (wt/v) sucrose and 0.37% (wt/v) Phytagel (Sigma).
For flowering time measurement, plants were grown on soil containing a 3 1 mixture of substrate and vermiculite in a temperature-controlled greenhouse environment with 16L 8D long-day and 8L:16D short-day cycle.
27a Later, they found the enantiomeric excess of this reaction could be dramatically improved to 99% ee when a solution of NFSI was slowly added to the mixture of substrate and catalyst.
After applying with enzyme conjugate for 10 min, the slides were incubated with the mixture of substrate buffer, chromogen solution and 0.6% hydrogen peroxide for HRP and monitored under a microscope.
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Presence of water in the mixture of substrates has a beneficial effect in maintaining a high catalyst activity.
However, the mixture of substrates inside the pitchers plays a key role in the success of the combined system.
In a test arrangement without separating and recycling o-cresol, that kind of state can be created artificially by placing it in the mixture of substrates and selecting conditions for the synthesis process to ensure that the amount of this component before and after passing through the catalyst layer is the same.
Consequently, the most general way to compare different substrates is the catalytic efficiency (Vmax/Km), which also reflects the ratio of conversion when a mixture of substrates is offered to an enzyme.
Instead, it may be preferable to screen under conditions that contain a complex mixture of substrates, ideally mimicking as closely as possible the substrate milieu in which therapeutic compounds are eventually intended to act.
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