Additional file 1 of Production of (R)-mandelic acid from styrene, L-phenylalanine, glycerol, or glucose via cascade biotransformations
Creators: name: Lukito, Benedict Ryan
name: Zilong Wang
name: Balaji Sundara Sekar
name: Li, Zhi
Publisher: name: figshare
Year: 2021
Subjects: subject: Organic Chemistry
subject: FOS: Chemical sciences
subject: FOS: Chemical sciences

description: Additional file 1: Table S1. Strains used in this study. Table S2. Enzymes used in this study. Fig. S1. Standard calibration curve for GC analysis of styrene. Fig. S2. Standard calibration curve for GC analysis of (S)-styrene oxide. Fig. S3. GC chromatogram of styrene (standard) with benzyl alcohol as internal standard (I.S.). Fig. S4. GC chromatogram of (S)-styrene oxide (standard) with benzyl alcohol as internal standard (I.S.). Fig. S5. Standard calibration curve for HPLC analysis of L-Phe. Fig. S6. Standard calibration curve for HPLC analysis of (R)-PED. Fig. S7. Standard calibration curve for HPLC analysis of (R)-MA. Fig. S8. HPLC chromatogram of L-Phe (standard) with benzyl alcohol as internal standard (I.S.). Fig. S9. HPLC chromatogram of (R)-PED (standard) with benzyl alcohol as internal standard (I.S.). Fig. S10. HPLC chromatogram of (R)-MA (standard) with benzyl alcohol as internal standard (I.S.). Fig. S11. HPLC chromatogram of (R)-MA produced during the biotransformation of (R)-PED (20 mM) with resting cells of E. coli (AldO) cells (15 g cdw/L) in KP buffer (200 mM, pH 8.0) at 30 °C for 24 h. Benzyl alcohol was used as internal standard (I.S.). Fig. S12. HPLC chromatogram of (R)-MA produced during the biotransformation of styrene (20 mM) with resting cells of E. coli (S2A) cells (15 g cdw/L) in a mixture of KP buffer (200 mM, pH 8.0) containing 0.5% glucose and n-hexadecane (1:1, v/v) at 30 °C for 24 h. Benzyl alcohol was used as internal standard (I.S.). Fig. S13. HPLC chromatogram of (R)-MA produced during the biotransformation of L-Phe (15 mM) with resting cells of E. coli (P2S2A) cells (15 g cdw/L) in a mixture of KP buffer (200 mM, pH 8.0) containing 0.5% glucose and n-hexadecane (1:1, v/v) at 30 °C for 24 h. Benzyl alcohol was used as internal standard (I.S.). Fig. S14. HPLC chromatogram of (R)-MA formed after the biotransformation from glycerol with growing cells of E. coli NST74 (P2S2A). Reaction was performed in a mixture of KP buffer (200 mM, pH 8.0) and n-hexadecane (2:1, v/v) at 25 °C for 24 h. Benzyl alcohol was used as internal standard (I.S.). Fig. S15. HPLC chromatogram of (R)-MA produced from glycerol via coupling E. coli NST74-Phe cells for the production of L-Phe and E. coli (P2S2A) cells (15 g cdw/L) for the production of (R)-MA in a reaction mixture containing KP buffer (200 mM, pH 8.0, 0.5% glucose) and n-hexadecane (1:1, v/v) at 30 °C for 24 h. Benzyl alcohol was used as internal standard (I.S.). Fig. S16. Chiral HPLC chromatograms of (R)-MA. a) Racemic MA standard, b) (R)-MA standard, c) Isolated (R)-MA products from biotransformation of L-Phe to (R)-MA with E. coli (P2S2A) cells. Fig S17. NMR spectrum of (R)-MA prepared from biotransformation of L-Phe with E. coli (P2S2A) cells. a) 1H NMR spectrum (400 MHz, CD3OD), δ 7.47 (dd, J = 15.1, 7.2 Hz, 2H), 7.31 (ddd, J = 10.7, 9.0, 5.1 Hz, 3H), 5.19 (s, 1H). b) 13C NMR spectrum (101 MHz, CD3OD), δ 176.27, 140.75, 129.53, 129.33, 127.96, 74.21. Fig. S18. Time course of biotransformation of glucose to (R)-MA via coupling E. coli NST74-Phe and E. coli (P2S2A) cells in aqueous-n-hexadecane two-phase system. Fig. S19. SDS-PAGE analysis of total intracellular protein of E. coli cells used for the bioproduction of (R)-MA; Lane A: E. coli (AldO); B: E. coli (S2A); C: E. coli (P2S2A).