Magnesium sulphate

4‐(Pentafluorosulfanyl)aniline was obtained from Fluorochem and 5‐methyl‐4‐isoxazolecarbonyl chloride was purchased from Apollo Scientific. Magnesium sulphate and sodium bicarbonate were obtained from Fisher Scientific. Preparative TLC plates were obtained from Analtech. Solvents and reagents were purchased from commercial suppliers and were used without purification. All reactions were performed in a fume hood. NMR spectra were recorded on Varian 500 MHz or 400 MHz spectrometers and chemical shifts are reported in ppm, usually referenced to TMS as an internal standard. LCMS measurements were performed on a Shimadzu LCMS‐2020 equipped with a Gemini® 5 μm C18 110 Å column and percentage purity measurements were run over 30 minutes in water/acetonitrile with 0.1 % formic acid (5 min at 5 %, 5 %–95 % over 20 min, 5 min at 95 %) with the UV detector at 254 nm. Mass spectrometry: ESI mass spectra were obtained using a Bruker Daltonics Apex III, using Apollo ESI as the ESI source. For EI mass spectra, a Fissions VG Autospec instrument was used at 70 eV. All analyses were run by Dr. Alla K. Abdul Sada. Analyses are for the molecular ion peak [M]⁺ and are given in m/z, mass to charge ratio. Melting points were determined using a Stanford Research Systems Optimelt and are uncorrected.

All chemicals were used as supplied. Ethyl acetate, hexane, methanol, dichloromethane, tetrahydrofuran, and magnesium sulphate were all purchased from Fisher Scientific at laboratory reagent grade. Deuterated chloroform (99.8 atom% D), dimethyl sulfoxide-d6 (99.9 atom% D), vinyl acetate (97%), potassium ethyl xanthate (96%), 2-(methyl bromopropionate) (98%), bismuth(iii) oxide powder (99.999% trace metals basis) hydrazine hydrate 50–60%, phosphate buffered saline tablet pH 7.4 (NaCl = 0.137 M and KCl = 0.0027 M at 25 °C), were purchased from Sigma Aldrich. Dialysis membrane (MWCO = 1000 Da/MWCO: 300–500 Da) was purchased from Spectra/Por.

Amitriptyline hydrochloride (AMI-HCl), Brij™ O20, dichloromethane (DCM), anhydrous acetone, and sodium carbonate were obtained from Sigma-Aldrich (UK). Sodium dihydrogen phosphate, sodium hydrogen phosphate, magnesium sulphate, and dithiothreitol (DTT) were supplied by Fisher Scientific (UK). Phosphate-buffered saline (pH 7.3 ± 0.2 at 25 °C) was prepared using Dulbecco A tablets supplied by Oxoid (UK). Octyl salicylate (OSAL), PG, IPM, IPA, and 1-octanol were obtained from Sigma-Aldrich (UK). High-performance liquid chromatography (HPLC) grade water, HPLC grade acetonitrile, HPLC grade methanol, liquid chromatography/mass spectrometry (LC-MS) grade acetonitrile, LC-MS grade 0.1% trifluoroacetic acid (TFA) solution, limonene (LIM), ethyl acetate (EtAc), and TFA were supplied by Fisher Scientific (UK). Chloroform-d (CDCl₃) were obtained from Cambridge Isotope Laboratories Inc. (UK). Propylene glycol monolaurate (PGML) Type II and TC were gifts from Gattefossé (St. Priest, France). All chemicals, unless otherwise stated, were ≥ 99% purity.

All chemicals were used as supplied. Ethyl acetate, hexane, methanol, Petroleum ether 40–60 °C dichloromethane and magnesium sulphate were all purchased from Fisher Scientific at laboratory reagent grade. Deuterated chloroform (99.8 atom %D), dimethyl sulfoxide-d6 (99.9 atom %D), vinyl acetate (97.0%), 4,4′-azo-bis(4-cyanovaleric acid) (≥80.0%), 2,2′-azo-bis(2-methylpropionitrile) (98%), potassium ethyl xanthate (96%), 2-(methyl bromopropionate) (98%), aqueous hydrazine hydrate solution (50–60%), PBS buffer (preformulated tablets, yielding 0.01 M phosphate buffer, 0.0027 M potassium chloride and 0.137 M sodium chloride, pH 7.4), N-Ethyl-N′-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC.HCl, >99.0%), N-hydroxy succinimide (NHS, 98%), imidazole (>99%), β-nicotinamide adenine dinucleotide, reduced disodium salt hydrate (NADH 97%), sodium pyruvate (ReagentPlus > 99%), polyethylene glycol (BioUltra 4000), poly(vinyl alcohol) (MW 9–10 kDa, 80% hydrolysed) and SealPlate films were purchased from Sigma Aldrich. L–lactate dehydrogenase was purchased from Roche. MilliQ water (18.2 mΩ).
Protein expression and purification are described in the Supporting Information.

The following analytical grade reagents were obtained from commercial suppliers. Tetrahydrofuran, dichloromethane, sodium bicarbonate, magnesium sulphate, methanol, ethyl acetate and hydrochloric acid were obtained from Fisher Scientific (Loughborough, UK). Acetone was acquired from Sigma-Aldrich (Dorset, UK). 3-Fluoroaniline was obtained from Fluorochem (Derbyshire, UK). Benzyl alcohol was obtained from Acros Organics (Geel, Belgium). 1-Methyl-2-pyrrolidinone, isobutyl chloroformate, n-(tert-butoxycarbonyl)-d-alanine and citric acid were obtained from Alfa Aesar (Lancashire, UK). n-Methylmorpholine was obtained from Lancaster Synthesis (Lancaster, UK). Benzyl-α-d-mannopyranoside was obtained from Santa Cruz Biotechnology (Heidelberg, Germany). Listeria enrichment broth base CM0862 was obtained from Oxoid Limited (Basingstoke, UK).