Journal: bioRxiv

Article Title: Mechanistic basis for the emergence of EPS1 as a catalyst in plant salicylic acid biosynthesis

doi: 10.1101/2021.08.21.457228

Figure Lengend Snippet: A vigorously stirred solution of phenol S1 (500 mg, 2.58 mmol) in DMF (26.0 mL) at 0 °C was treated with NaH (60% in mineral oil) (124 mg, 3.10 mmol). The reaction mixture was warmed to 23 °C, stirred for 30 min, and then treated with bromide S2 (0.43 mL, 3.9 mmol). The reaction mixture was stirred at 23 °C for 20 h. After 20 h, saturated aqueous NH 4 Cl (50 mL), H 2 O (100 mL), and EtOAc (150 mL) were added to the reaction mixture. The organic layer was separated and the aqueous layer was extracted with EtOAc (2 × 50 mL). The combined organic extracts were dried over Na 2 SO 4 and concentrated on a rotary evaporator. Flash chromatography (SiO 2 , 5–10% EtOAc/hexanes) provided S3 as a colorless film (606 mg, 84%): 1 H NMR (CDCl 3 , 400 MHz) δ 7.63 (d, J = 7.7 Hz, 1H), 7.51 (s, 1H), 7.33 (t, J = 8.0 Hz, 1H), 7.09 (dd, J = 8.4, 2.7 Hz, 1H), 4.65 (s, 2H), 4.27 (q, J = 7.1 Hz, 2H), 1.58 (s, 9H), 1.30 (t, J = 7.2 Hz, 2H); 13 C NMR (CDCl 3 , 100 MHz) δ 168.8, 165.4, 157.8, 133.6, 129.5, 123.0, 119.5, 115.0, 81.3, 65.6, 61.6, 28.3, 14.3; IR (film) ν max 2978, 2932, 1758, 1711, 1586, 1486, 1440, 1368, 1294, 1193, 1163, 1105, 1083, 849, 808, 681 cm −1 ; HRMS (DART-TOF) m/z 280.1314 (C 15 H 20 O 5 + H + requires 281.1389).

Article Snippet: Column chromatography was conducted using Silicycle SiliaFlash P60 SiO 2 (40–63 μm).

Techniques: Chromatography