Biocatalytic bis-C-alkylation of phenolics using one-pot cascades with promiscuous C-glycosyltransferase and prenyltransferase

doi:10.5246/jcps.2018.04.025

Abstract

Abstract:

C-glycosylation and C-prenylation are two important C-C-bond forming reactions for preparation, diversification and structural modification of natural/unnatural products with pharmacological activities. Here, we described unprecedented enzymaticcascades to C-glycosylate/prenylate different acyl resorcinol derivatives in stepwise, one-pot reactions by combining two promiscuous enzymes, MiCGT, a C-glycosyltransferase, and AtaPT, a prenyltransferase. Five novel bis-C-alkylated products were obtained and structurally identified by MS and NMR spectroscopic data as well as comparison with the literature. This study provided a potential synthetic strategy for synthesizing structurally novel and diverse compounds bearing both C-glycosyl and C-prenyl moieties by a two-step, enzymatic bis-C-alkylation.

Key words: Bis-C-alkylation, Enzymatic glycosylation, Enzymatic prenylation, Enzyme cascades

CLC Number:

R917

Supporting:

Figures S1–S8. HPLC-DAD/ESI-MS analysis of C-glycosylated and C-prenylated products.

Figures S9–S14. HPLC-DAD/ESI-MS analysis of C-prenylated and C-glycosylated products.

Figure S15. HPLC analysis of C-glycosyl and geranyl products of 1.

Figures S16–S20. HR-ESI-MS spectra of C-glycosyl/prenyl products.

Figures S21–S30. ¹H and ¹³C NMR spectra of C-glycosyl/prenyl products.

Figure S1. HPLC-DAD/ESI-MS analysis of C-glycosylated and C-prenylated product 1GA. The C-glycosylation of 1 was performed by MiCGT and UDP-Glc; after 12 h, AtaPT and DMAPP were added for C-prenylation of 1G. (a) HPLC chromatogram and UV spectra of enzyme products 1G and 1GA; (b) Typical negative ion MS spectrum for 1GA;(c) Typical negative ion MS² spectrum for 1GA. The product 1G is a C-glycosylated compound.

Figure S2. HPLC-DAD/ESI-MS analysis of C-glycosylated and C-prenylated product 1GB. The C-glycosylation of 1 was performed by MiCGT and UDP-Glc; after 12 h, AtaPT and GPP were added for C-prenylation of 1G. (a) HPLC chromatogram and UV spectra of enzyme products 1G and 1GB; (b) Typical negative ion MS spectrum for 1GB;(c) Typical negative ion MS² spectrum for 1GB. The product 1G is a C-glycosylated compound.

Figure S3. HPLC-DAD/ESI-MS analysis of C-glycosylated and C-prenylated product 1GC. The C-glycosylation of 1 was performed by MiCGT and UDP-Glc; after 12 h, AtaPT and FPP were added for C-prenylation of 1G. (a) HPLC chromatogram and UV spectra of enzyme products 1G and 1GC; (b) Typical negative ion MS spectrum for 1GC. The product 1G is a C-glycosylated compound.

Figure S4. HPLC-DAD/ESI-MS analysis of C-glycosylated and C-prenylated product 2GA. The C-glycosylation of 2 was performed by MiCGT and UDP-Glc; after 12 h, AtaPT and DMAPP were added for C-prenylation of 2G. (a) HPLC chromatogram and UV spectra of enzyme products 2G and 2GA; (b) Typical negative ion MS spectrum for 2GA;(c) Typical negative ion MS² spectrum for 2GA. The product 2G is a C-glycosylated compound.

Figure S5. HPLC-DAD/ESI-MS analysis of C-glycosylated and C-prenylated product 3GB. The C-glycosylation of 3 was performed by MiCGT and UDP-Glc; after 12 h, AtaPT and GPP were added for C-prenylation of 3G. (a) HPLC chromatogram and UV spectra of enzyme products 3G and 3GB; (b) Typical negative ion MS spectrum for 3GB;(c) Typical negative ion MS² spectrum for 3GB. The product 3G is a C-glycosylated compound.

Figure S6. HPLC-DAD/ESI-MS analysis of C-glycosylated and C-prenylated product 4GB. The C-glycosylation of 4 was performed by MiCGT and UDP-Glc; after 12 h, AtaPT and GPP were added for C-prenylation of 4G. (a) HPLC chromatogram and UV spectra of enzyme products 4G and 4GB; (b) Typical negative ion MS spectrum for 4GB;(c) Typical negative ion MS² spectrum for 4GB. The product 4G is a C-glycosylated compound.

Figure S7. HPLC-DAD/ESI-MS analysis of C-glycosylated and C-prenylated product 4GC. The C-glycosylation of 4 was performed by MiCGT and UDP-Glc; after 12 h, AtaPT and FPP were added for C-prenylation of 4G. (a) HPLC chromatogram and UV spectra of enzyme products 4G and 4GC; (b) Typical negative ion MS spectrum for 4GC. The product 4G is a C-glycosylated compound.

Figure S8. HPLC-DAD/ESI-MS analysis of C-glycosylated and C-prenylated product 5GB. The C-glycosylation of 5 was performed by MiCGT and UDP-Glc; after 12 h, AtaPT and GPP were added for C-prenylation of 5G. (a) HPLC chromatogram and UV spectra of enzyme products 5G and 5GB; (b) Typical negative ion MS spectrum for 5GB;(c) Typical negative ion MS² spectrum for 5GB. The product 5G is a C-glycosylated compound.

Figure S9. HPLC-DAD/ESI-MS analysis of C-prenylated and C-glycosylated product 6AG. The C-prenylation of 6 was performed by AtaPT and DMAPP; after 12 h, MiCGT and UDP-Glc were added for C-glycosylation. (a) HPLC chromatogram and UV spectra of enzyme products 6G and 6AG; (b) Typical negative ion MS spectrum for 6AG;(c) Typical negative ion MS² spectrum for 6AG. The product 6G is a C-glycosylated compound.

Figure S10. HPLC-DAD/ESI-MS analysis of C-prenylated and C-glycosylated product 6BG. The C-prenylation of 6 was performed by AtaPT and GPP; after 12 h, MiCGT and UDP-Glc were added for C-glycosylation. (a) HPLC chromatogram and UV spectra of enzyme products 6G, 6BG and 6B; (b) Typical negative ion MS spectrum for 6BG;(c) Typical negative ion MS² spectrum for 6BG. The product 6G is a C-glycosylated compound. The product 6B is a C-prenylated compound.

Figure S11. HPLC-DAD/ESI-MS analysis of C-prenylated and C-glycosylated product 6CG. The C-prenylation of 6 was performed by AtaPT and FPP; after 12 h, MiCGT and UDP-Glc were added for C-glycosylation. (a) HPLC chromatogram and UV spectra of enzyme products 6CG, 6Cand 6C′; (b) Typical negative ion MS spectrum for 6CG;(c) Typical negative ion MS² spectrum for 6CG. The products 6C and 6C' are C-prenylated compounds.

Figure S12. HPLC-DAD/ESI-MS analysis of C-prenylated and C-glycosylated product 7GA. The C-prenylation of 7 was performed by AtaPT and DMAPP; after 12 h, MiCGT and UDP-Glc were added for C-glycosylation. (a) HPLC chromatogram and UV spectra of enzyme products 7G and 7GA; (b) Typical negative ion MS spectrum for 7GA; (c) Typical negative ion MS² spectrum for 7GA. The product 7G is a C-glycosylated compound.

Figure S13. HPLC-DAD/ESI-MS analysis of C-prenylated and C-glycosylated product 7BG. The C-prenylation of 7 was performed by AtaPT and GPP; after 12 h, MiCGT and UDP-Glc were added for C-glycosylation. (a) HPLC chromatogram and UV spectra of enzyme products 7G, 7BG and 7B; (b) Typical negative ion MS spectrum for 7BG; (c) Typical negative ion MS² spectrum for 7BG. The product 7G is a C-glycosylated compound. The product 7B is a C-prenylated compound.

Figure S14. HPLC-DAD/ESI-MS analysis of C-prenylated and C-glycosylated product 7CG. The C-prenylation of 7 was performed by AtaPT and FPP; after 12 h, MiCGT and UDP-glucose were added for C-glycosylation. (a) HPLC chromatogram and UV spectra of enzyme products 7G and 7CG; (b) Typical negative ion MS spectrum for 7CG. The product 7G is a C-glycosylated compound.

Figure S15. HPLC analysis of C-glycosyl and geranyl products of 1. The reaction was performed with AtaPT and GPP, and MiCGT and UDP-glucose in simultaneous mode.

Figure S16. HR-ESI-MS spectrum for 1GB.

Figure S17. HR-ESI-MS spectrum for 1GC.

Figure S18. HR-ESI-MS spectrum for 4GB.

Figure S19. HR-ESI-MS spectrum for 6BG.

Figure S20. HR-ESI-MS spectrum for 7BG.

Figure S21. ¹H NMR spectrum of 1GB (DMSO-d₆, 400 MHz).

Figure S22. ¹³C NMR spectrum of 1GB (DMSO-d₆, 150 MHz).

Figure S23. ¹H NMR spectrum of 1GC (DMSO-d₆, 400 MHz).

Figure S24. ¹³C NMR spectrum of 1GC (DMSO-d₆, 150 MHz).

Figure S25. ¹H NMR spectrum of 4GB (DMSO-d₆, 400 MHz).

Figure S26. ¹³C NMR spectrum of 4GB (DMSO-d₆, 150 MHz).

Figure S27. ¹H NMR spectrum of 6BG (DMSO-d₆, 400 MHz).

Figure S28. ¹³C NMR spectrum of 6BG (DMSO-d₆, 150 MHz).

Figure S29. ¹H NMR spectrum of 7BG (DMSO-d₆, 400 MHz).

Figure S30. ¹³C NMR spectrum of 7BG (DMSO-d₆, 150 MHz).

Dawei Chen, Lili Sun, Ridao Chen, Kebo Xie, Lin Yang, Jungui Dai. Biocatalytic bis-C-alkylation of phenolics using one-pot cascades with promiscuous C-glycosyltransferase and prenyltransferase[J]. Journal of Chinese Pharmaceutical Sciences, 2018, 27(4): 241-250.

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