Aryl and aliphatic terminal alkynes are also competent reaction partners; however, product distributions are highly variable, depending on the substrate.
Although a variety of cyclic allylic electrophiles were found to be competent reaction partners, only hydrodehalogenation was observed with an acyclic electrophile.
Both boronic acids and trifluoroborate salts were competent reaction partners, and products bearing interesting heterocyclic scaffolds could be prepared with good efficiency.
This is consistent with the copper(I) amide complexes being competent reaction intermediates in the modified Ullmann amination reaction and also represents the first experimental evidence of copper(I) amide aggregates reacting with aryl halides in the absence of any ancillary ligands.
To assess whether the cross-linked species, presumably 6-mercaptooctanoyl-H protein bound to LS via an S Fe bond to the auxiliary cluster, is catalytically and kinetically competent, LS reactions were conducted with peptide 1, which is not labeled with deuterium at C8.
Although kcat/ KM O2) is orders of magnitude slower than expected for a diffusional process (ca. 1 × 10 M–1 s–1), we could not dismiss the possibility that there was an unfavorable pre-equilibrium leading to a very small fraction of FIH being competent for reaction upon collision.
Interestingly, colon microsomes were competent in this reaction, showing ∼60% of the activity seen in SI microsomes in WT mice (Table 2).
Both aryl and aliphatic vinyl trifluoroborates are competent in the reaction; however, the scope of electrophiles was limited to aryl- and heteroaryl-substituted allylic alcohols.
While in several of the cases substrates indeed ranked in the top of the docking hit list (Supplementary Table 1, Supporting Information), it was often the case that the predicted docking pose was not competent for the reaction (Supplementary Figure 2, Supporting Information).
We then electroporated the resulting 4 independent ligation reactions into competent cells of strain SQ765 (genotype: MG1655, Δ hsdR) (Table 1) to generate four independent mutant plasmid libraries, each containing ∼10 clones.
50 µl of distilled water was then added to each reaction and competent JM109 E.coli cells (Promega: #L2001) were chemically transformed with 10 µl of the diluted ligation reaction as per standard methods and grown overnight at 37°C on LB (Luria broth) agar media supplemented with ampicillin using standard (X-Gal/IPTG) blue/white selection.
