C-糖基转移酶及异戊烯转移酶偶联催化酚类化合物的bis-C-烷基化

doi:10.5246/jcps.2018.04.025

中国药学(英文版) ›› 2018, Vol. 27 ›› Issue (4): 241-250.DOI: 10.5246/jcps.2018.04.025

C-糖基转移酶及异戊烯转移酶偶联催化酚类化合物的bis-C-烷基化

陈大伟^1#, 孙丽丽^2#, 陈日道¹, 解可波¹, 杨林², 戴均贵^1*

1. 中国医学科学院北京协和医学院药物研究所天然药物活性物质与功能国家重点实验室, 北京 100050
2. 中央民族大学生命与环境科学学院, 北京 100081

收稿日期:2018-02-01 修回日期:2018-03-05 出版日期:2018-04-30 发布日期:2018-03-11
通讯作者: Tel.: +86-010-63165195; Fax: +86-010-63017757, E-mail: 15116995486@163.com; jgdai@imm.ac.cn
基金资助:
National Natural Science Foundation of China (Grant Nos. 21572277, 81573317 and 81703369) and CAMS Innovation Fund for Medical Sciences (CIFMS-2016-I2M-3-012).

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

Dawei Chen^1#, Lili Sun^2#, Ridao Chen¹, Kebo Xie¹, Lin Yang^2*, Jungui Dai^1*

1. State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China
2. College of Life and Environmental Sciences, Minzu University of China, Beijing 100081, China

Received:2018-02-01 Revised:2018-03-05 Online:2018-04-30 Published:2018-03-11
Contact: Tel.: +86-010-63165195; Fax: +86-010-63017757, E-mail: 15116995486@163.com; jgdai@imm.ac.cn
Supported by:
National Natural Science Foundation of China (Grant Nos. 21572277, 81573317 and 81703369) and CAMS Innovation Fund for Medical Sciences (CIFMS-2016-I2M-3-012).

摘要/Abstract

摘要：

C-糖基化和C-异戊烯化是两个重要的C-C键形成反应,可用于具有药理活性天然/非天然产物的制备,多样化及结构修饰。本研究中,我们将具有杂泛性的C-糖基转移酶MiCGT和异戊烯基转移酶AtaPT偶联,一锅法对酰基间苯二酚类化合物分步进行C-糖基化/异戊烯基化修饰。通过MS和NMR数据分析及文献对比,我们获得了5个新的bis-C-烷基化产物。本研究通过偶联两步酶促bis-C-烷基化反应,为结构新颖及多样化且同时含有C-糖基和C-异戊烯基类化合物的合成提供了一种潜在策略。

关键词: Bis-C-烷基化, 酶法糖基化, 酶法异戊烯基化, 酶偶联

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

中图分类号:

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).

陈大伟, 孙丽丽, 陈日道, 解可波, 杨林, 戴均贵. C-糖基转移酶及异戊烯转移酶偶联催化酚类化合物的bis-C-烷基化[J]. 中国药学(英文版), 2018, 27(4): 241-250.

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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C-糖基转移酶及异戊烯转移酶偶联催化酚类化合物的bis-C-烷基化

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

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