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With hydrogenchloride; In water; at 100℃; for 6.0h;
(S)-7'-Chlorotryptophan, 3. Method A: (S)-N-acetyl-7'-chlorotryptophan was refluxed in 3 N HCl for 6 h to remove the acetyl group. Concentration of the reaction mixture gave x mg of (S)-7'-chlorotryptophan ( %).
With disodium hydrogenphosphate; Rhodococcus ruber alcohol dehydrogenase; flavin reductase PrnF; tryptophan 6-halogenase from Streptomyces albogriseolus RebH5 variant; NAD; flavin adenine dinucleotide; sodium chloride; In isopropyl alcohol; at 25℃; for 86.0h;pH 7.4;Enzymatic reaction;
Enzymatic chlorination was assayed in a total volume of 500lwhileshakinggentlyat25C.Substrate L -tryptophan wasadded to a final concentration of 5 m M together with 1 m MNAD, 0.01 m M FAD, 1 unit ml 1RR-ADH, 2.5 units ml 1flavin reductase PrnF, 5% (v/v) isopropyl alcohol, 10 m MNa 2 HPO 4 , pH 7.4, and 30 m M NaCl. Purified halogenase wasadded to a final concentration of 89M . Reactions were per-formed as triplicates, and reaction progress was monitored byHPLC-MS or analytical HPLC. Aliquots of 30lweretakenfrom the reaction mixture at different time points andquenched by adding an equal volume of methanol. The mixturewas centrifuged (12,000 g, 10 min), and the supernatant wasanalyzed via HPLC-MS or analytical HPLC.
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