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m is just not minimal by the Affiliation rate frequent for the formation of the NagA-substrate elaborate. These success (along with the improved fee of reaction of compound 6 relative to compound 1) are only consistent with the summary that NagA is proscribed by either the development or breakdown with the tetrahedral intermediate.

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Reconstitution of the apo-enzymes was carried out overnight at 4 °C prior to removing of unbound metal by passage through a PD-ten column. The PD-10 column was pre-treated with dipicolinate to remove traces of unbound steel after which you can washed with five column volumes of steel cost-free HEPES buffer, pH 8.0. Immediately after elution, the focus of your metallic-reconstituted enzyme was determined by UV absorbance and also the steel articles with the samples based on ICP-MS.

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