Additional file 1 of One-pot synthesis of (R)- and (S)-phenylglycinol from bio-based l-phenylalanine by an artificial biocatalytic cascade
Creators: name: Zhang, Jiandong
name: Qi, Ning
name: Gao, Lili
name: Li, Jing
name: Zhang, Chaofeng
name: Chang, Honghong
Publisher: name: figshare
Year: 2021
Subjects: subject: Organic Chemistry
subject: FOS: Chemical sciences

description: Additional file 1: Table S1. Primers used in this study. Table S2. Recombinant strains constructed in this study. Table S3. The specific activity of enzymes used in this study. Figure S1. Recombinant plasmid constructed. Figure S2. Recombinant E. coli cells co-expression of multiple enzymes. Figure S3. SDS-PAGE of the cell-free extracts of recombinant E. coli cells co-expression of multiple enzymes. A: lane M:marker, lane 1, 2: E. coli (DEA), lane 3, 4: E. coli (CGS); B: laneM:marker, lane 1, 2: E. coli (REG); C: laneM:marker, lane 1, 2: E. coli (DFP); D: laneM:marker, lane1: E. coli (CFP); E: laneM:marker, lane 1, 2: E. coli (RFP), lane 3: E. coli (DES); F: lane M:marker, lane 1, 2: E. coli (RGA); G: lane M:marker, lane 1: E. coli (CAS). Figure S4. Achiral GC chromatograms of 2-HAP. A: 2-HAP standard. B: 2-HAP produced by conversion of L-phenylalanine (10 mM) with resting cells of E. coli (RFP-DEA-CGS) (15 g cdw/L) at 3 h. C: 2-HAP produced by conversion of L-phenylalanine (20 mM) with resting cells of E. coli (RFP-DEA-CGS) (25 g cdw/L) at 9 h. D: 2-HAP produced by conversion of L-phenylalanine (50 mM) with resting cells of E. coli (RFP-DEA-CGS) (25 g cdw/L) at 12 h. IS: Internal standard (n-dodecane), 4.05 min; 2-HAP: 2-hydroxyacetophenone, 4.91 min. Figure S5. Effect of different concentration of RMBA on E. coli (RFP-DEA-CGS) cells for conversion of L-PA to 2-HAP. Figure S6. Achiral GC chromatograms of (S)-phenylglycinol. A: (S)-phenylglycinol standard. B: (S)-phenylglycinol produced by conversion of L-phenylalanine (10 mM) with resting cells of E. coli (RFP-DEA-CGS) (20 g cdw/L) and E. coli (MVTA) (15 g cdw/L) at 12 h. C: (S)-phenylglycinol produced by conversion of L-phenylalanine (20 mM) with resting cells of E. coli (RFPDEA-CGS) (20 g cdw/L) and E. coli (MVTA) (15 g cdw/L) at 12 h. IS: Internal standard (n-dodecane), 4.05 min; RMBA: (R)-(+)-1-phenylethylamine, 2.62 min; AP: acetophenone, 2.78 min; Product (S)-phenylglycinol, 6.05 min. Figure S7. Achiral GC chromatograms of (R)-phenylglycinol. A: (R)-phenylglycinol standard. B: (R)-phenylglycinol produced by conversion of L-phenylalanine (10 mM) with resting cells of E. coli (RFP-DEA-CGS) (15 g cdw/L) and E. coli (EB-DGA) (20 g cdw/L) at 9 h. C: (R)-phenylglycinol produced by conversion of L-phenylalanine (20 mM) with resting cells of E. coli (RFP-DEA-CGS) (15 g cdw/L) and E. coli (EB-DGA) (20 g cdw/L) at 12 h. IS: Internal standard (n-dodecane), 4.05 min; (R)-phenylglycinol, 6.05 min. Figure S8. Chiral GC chromatograms of phenylglycinol. A: (±)- phenylglycinol standard. B: (R)-phenylglycinol standard. C: (S)-phenylglycinol standard. D: (R)-phenylglycinol produced by conversion of L-phenylalanine (20 mM) with resting cells of E . coli (RFP-DEA-CGS) (15 g cdw/L) and E. coli (EB-DGA) (20 g cdw/L) at 12 h. E: (S)-phenylglycinol produced by conversion of L-phenylalanine (20 mM) with resting cells of E. coli (RFP-DEA-CGS) (20 g cdw/L) and E. coli (MVTA) (15 g cdw/L) at 9 h. (R)-phenylglycinol, 28.7 min; (S)-phenylglycinol, 29.3 min. Figure S9. Chiral GC chromatograms of phenylglycinol. A: (±)- phenylglycinol standard. B: (R)-phenylglycinol standard. C: (S)-phenylglycinol standard. D: Preparation of (R)-phenylglycinol from L-phenylalanine. E: Preparation of (S)-phenylglycinol from L-phenylalanine. (R)-phenylglycinol, 28.7 min; (S)-phenylglycinol, 29.3 min. Figure S10. 1H NMR spectra analysis of 7.