3 pyridinemethanol

3-pyridinemethanol, potassium tert-butoxide, toluene, dichloromethane, ethylmalonic acid, 2,2-dimethylmalonic acid, methylsuccinic acid, glutaric acid, n-adipic acid, n-suberic acid, n-sebacic acid, n-octanoylglycine, n-decanoylglycine, glycine methylester hydrochloride, 2-ethylhexanoic acid, 2-ethyloctanoic acid, 4-ethyloctanoic acid, tetrahydrofurane, carbonyldiimidazole, 2-mercapto-ethanol, 2-(N-morpholino) ethanesulfonic acid (MES) monohydrate, and coenzyme A were from Sigma-Aldrich (now Merck). ethylmalonic acid (methyl-d3) was from Cambridge Isotope Laboratories. Methanol, acetonitrile, isopropanol (MS-grade), formic acid, and ammonium formate (all MS-grade) were from Biosolve. n-Hexane was from VWR chemicals. Ethylmalonyl-CoA was from CoALA Biosciences. Hydrochloric acid (37%) was from Merck. 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride (EDAC) was from Thermo Fisher Scientific.

Dicyandiamide (DCN, 99%), calcium chloride (CaCl₂, >99%), 3-Pyridinemethanol (3PM, 98%), 3-Pyridinecarboxaldehyde (3PC, 98%), and Vitamin B3 (VB3, ≥98%) were purchased from Sigma-Aldrich (Darmstadt, Germany). Formic acid (HCO₂H, 98%) was acquired from Fluka (Darmstadt, Germany). Methanol (CH₃OH, 99.6%) and sodium alginate were purchased from VWR^TM (Arnhem, The Netherlands). All the reagents were used without further purification. Ultra-pure water (18 MΩ cm⁻¹) was obtained from a Millipore Milli-Q water purification system (Darmstadt, Germany).

We adapted a method described by [37 (link)]. After tert-butyl methyl ether extraction, the organic layer was collected and passed through sodium sulfate drying cartridges (Agilent Technologies, Anhydrous Sodium sulf LRC 1 gm, 12131033) before being evaporated to dryness under nitrogen. The residue was dissolved in 1 ml dichloromethane (Sigma–Aldrich) and 300 µl of freshly prepared ‘picolinyl reagent' [2 : 1 v : v 3-pyridinemethanol (Sigma–Aldrich)/potassium tert-butoxide (Sigma–Aldrich)] was added for reaction in a stoppered glass tube for 30 min at 37°C. After cooling to room temperature, two steps of n-hexane extraction were carried out (as described above). Pooled organic fractions were dried through sodium sulfate drying cartridges and evaporated to dryness under nitrogen. The residue was reconstituted in 100 µl of hexane, and analyzed by GC–MS.