Microorganisms for Ginsenosides Biosynthesis: Recent Progress, Challenges, and Perspectives
Abstract
:1. Introduction
2. Ginsenosides: Classification and Cell Biological Mechanism in Anticancer Activities
3. Endophytes as Novel Biological Source of Ginsenosides
3.1. Bioproduction of Ginsenosides by Native Endophytes
3.2. Biotransformation of Major to Minor Ginsenosides by Endophytes
4. Ginsenoside Biosynthesis in Engineered Microorganisms
4.1. Ginsenoside Biosynthesis in Engineered Bacteria
4.2. Ginsenoside Production in Engineered Yeasts
Strains | Genes or Related Gene Cassettes | Products | Titer (mg/L) | Cultivation Condition | Major Media | Carbon Source | References |
---|---|---|---|---|---|---|---|
Saccharomyces cerevisiae | |||||||
ZD-PPD-018 | tHMG1, AtCPR1, SynPgPPDS, ERG20, ERG1, ERG9 | PPD | 1189 | Fed-batch | SD | Glucose | [79] |
DM | 1548 | ||||||
D20RH18 | PgDDS, synPgPPDS, ATR2.1, tHMG1, ERG20, PgERG1, ERG9, UGTPg45 | Rh2 | 1.45 μmol/g DCW | Shake-flask | YPD | Glucose | [74] |
D20RG1 | PgDDS, synPgPPDS, ATR2.1, tHMG1, ERG20, PgERG1, ERG9, UGTPg45, UGTPg29 | Rh3 | 3.49 μmol/g DCW | ||||
ZW-Rh1-20 | ERG20, PgERG1, ERG9, tHMG1, CYP716A53v2, PgCPR1, UGTPg100 | Rh1 | 98.2 | Shake-flask | SC | Glucose | [71] |
PPT | 3.5 | ||||||
PPD | 43.4 | ||||||
DM | 8.8 | ||||||
ZW-F1-17 | ERG20, PgERG1, ERG9, tHMG1, CYP716A53v2, PgCPR1, UGTPg1 | F1 | 42.1 | ||||
PPT | 13.9 | ||||||
CK | 7.5 | ||||||
PPD | 49.2 | ||||||
DM | 3.5 | ||||||
WLT-MVA5 | DS, PPDS-ATR1, ERG1, tHMG1, ERG9, ERG20, ERG10, ERG13, ERG12, ERG8, ERG19, IDI1, NCP1, ACSseL641P | PPD | 8090 | Fed-batch | YNBD | Glucose/ Ethanol | [80] |
Y1CSH | HAC1, IDI1, ERG20, ERG9, ERG1, ERG7, synDS-GFP, tHMG1, synPgUGT74AE2 | 3β-O-Glc-DM | 2400 | Fed-batch | YPD | Glucose | [72] |
Y2CSH | HAC1, IDI1, ERG20, ERG9, ERG1, ERG7, synDS-GFP, tHMG1, synUGTPg1 | 20S-O-Glc-DM | 5600 | ||||
PPD-A3-sgRNA4 | PgDS and PgPPDS, PgCPR, tHMGR1, ERG1m, ∆ ERG7 | PPD | 294.5 | Shake-flask | YPD | Glucose | [71] |
Rg1-02 | CYP716A53v2, PgUGT71A54, PgURT94, RHM | Rg2 | 1300 | Fed-batch | Synthetic | Glucose | [75] |
Re-01 | CYP716A53v2, PgUGT71A53, PgUGT71A54, PgURT94, RHM | Re | 3600 | ||||
CPX113436PPXP-ADH2 | ERG10, ERG13, tHMG1, ERG12, ERG8, ERG19, IDI1, ERG20, ERG9, ERG1, ERG7, PgDS, PgPPDS, PgCPR, ADH2, (Pex11p, Pex34p, and Atg36p) | PPD | 4.1 | Shake-flask | YPDO | Glucose and Ethanol | [81] |
BY-V | ERG10, ERG13, tHMG1, ERG12, ERG8, IDI1, MVD1, ERG20, ERG9, ERG1, PgDDS, AtCPR1, PgPPDS, INO2, ∆LPP1, ∆ERG7 | PPD | 1550 | Shake-flask | YPD | Sugarcane molasses | [78] |
158,800 | Fed-batch | ||||||
YFR | tHMG1, IDI1, ERG20, ERG9, ERG1, DS-GFP, PGM1, PGM2, INO2, ERG7, ERG1, PgUGT74AE2, UGTPg1, PP-DS, ATR2 | F2 | 21.0 | Shake-flask | YPD | Glucose | [73] |
YSR | tHMG1, IDI1, ERG20, ERG9, ERG1, DS-GFP, UGTPg1, PGM1, PGM2, INO2, ERG7, ERG1, M7 (ΔUGT74AC1) | 3β-20S-Di-O-Glc-DM | 346.1 | Glucose | |||
2600 | Fed-batch | ||||||
WEA | tHMG1, ERG10, ERG13,IDI, ERG20, ERG9, ERG1, UGD1, AeBAS1, AtATR2, AeCYP716A354, AeCSLM1, AeUGT74AG6 | Chikusetsusaponin IVa | NR | Shake-flask | SD | Glucose, Galactose | [82] |
tHMG1, ERG10, ERG13,IDI, ERG20, ERG9, ERG1, UGD1, AeBAS1, AtATR2, AeCYP716A354, AeCSLM1, AeUGT73CB3 | Zingibroside R1 | NR | |||||
ZY-M7(4)E1 PUA | ERG20, ERG1, ERG9, tHMG1, M7-1, ∆EGH1, PGM1, UGP1, PgPPDS-AtCPR2 | Rh2 | 300 | Fed-batch | SC | Glucose | [83] |
Yarrowia lipolytica | |||||||
Y14 | ΔLUL, XYL1, XYL2, ylXKS, DS, PPDS-linker-ATR1, tHMG1, ERG9, ERG20, TKL, TAL, TX | PPD | 300.63 | Fed-batch | YPD or YPX | Xylose | [84] |
167.17 | Glucose | ||||||
YL-MVA-CK | tHMG1, ERG9, ERG20, OpDS, PPDS-linker2-ATR1, UGT1 | CK | 161.8 | Fed-batch | YPD | Glucose | [85] |
Pichia pastoris | |||||||
KDPEP | PgDDS-L3-PDZlig and ERG1-ER/kPDZ with p-[PgDDS-PDZlig]/[ERG1-PDZ] | DM | 0.10 mg/g DCW | Shake-flask | YPD | Glucose, methanol | [86] |
5. Challenges and Future Perspectives
6. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- ** novel cell-factories in non-conventional yeasts. Biotechnol. Adv. 2021, 47, 107695. [Google Scholar]
- Yi, X.; Alper, H.S. Considering strain variation and non-type strains for yeast metabolic engineering applications. Life 2022, 12, 510. [Google Scholar] [CrossRef]
- Li, Y.; Wang, J.; Li, L.; Song, W.; Li, M.; Hua, X.; Wang, Y.; Yuan, J.; Xue, Z. Natural products of pentacyclic triterpenoids: From discovery to heterologous biosynthesis. Nat. Prod. Rep. 2022. [Google Scholar] [CrossRef]
- Luo, Y.; Jiang, Y.; Chen, L.; Li, C.; Wang, Y. Applications of protein engineering in the microbial synthesis of plant triterpenoids. Synth. Syst. Biotechnol. 2022, 8, 20–32. [Google Scholar] [CrossRef]
- Lim, S.H.; Baek, J.I.; Jeon, B.M.; Seo, J.W.; Kim, M.S.; Byun, J.Y.; Park, S.H.; Kim, S.J.; Lee, J.Y.; Lee, J.H.; et al. CRISPRi-Guided metabolic flux engineering for enhanced protopanaxadiol production in Saccharomyces cerevisiae. Int. J. Mol. Sci. 2021, 22, 11836. [Google Scholar] [CrossRef]
- Yan, X.; Fan, Y.; Wei, W.; Wang, P.; Liu, Q.; Wei, Y.; Zhang, L.; Zhao, G.; Yue, J.; Zhou, Z. Production of bioactive ginsenoside compound K in metabolically engineered yeast. Cell Res. 2014, 24, 770–773. [Google Scholar] [CrossRef]
- Wei, W.; Wang, P.; Wei, Y.; Liu, Q.; Yang, C.; Zhao, G.; Yue, J.; Yan, X.; Zhou, Z. Characterization of Panax ginseng UDP-Glycosyltransferases catalyzing protopanaxatriol and biosyntheses of bioactive ginsenosides F1 and Rh1 in metabolically engineered yeasts. Mol. Plant. 2015, 8, 1412–1424. [Google Scholar] [CrossRef] [PubMed]
- Hu, Z.F.; Gu, A.D.; Liang, L.; Li, Y.; Gong, T.; Chen, J.J.; Chen, T.J.; Yang, J.L.; Zhu, P. Construction and optimization of microbial cell factories for sustainable production of bioactive dammarenediol-II glucosides. Green. Chem. 2019, 21, 3286–3299. [Google Scholar] [CrossRef]
- Jiang, F.; Zhou, C.; Li, Y.; Deng, H.; Gong, T.; Chen, J.; Chen, T.; Yang, J.; Zhu, P. Metabolic engineering of yeasts for green and sustainable production of bioactive ginsenosides F2 and 3β,20S-Di-O-Glc-DM. Acta Pharm. Sin. B 2022, 12, 3167–3176. [Google Scholar] [CrossRef] [PubMed]
- Wang, P.; Wei, Y.; Fan, Y.; Liu, Q.; Wei, W.; Yang, C.; Zhang, L.; Zhao, G.; Yue, J.; Yan, X.; et al. Production of bioactive ginsenosides Rh2 and Rg3 by metabolically engineered yeasts. Metab. Eng. 2015, 29, 97–105. [Google Scholar] [CrossRef]
- Li, C.; Yan, X.; Xu, Z.; Wang, Y.; Shen, X.; Zhang, L.; Zhou, Z.; Wang, P. Pathway elucidation of bioactive rhamnosylated ginsenosides in Panax ginseng and their de novo high-level production by engineered Saccharomyces cerevisiae. Commun. Biol. 2022, 5, 775. [Google Scholar] [CrossRef]
- Dai, L.; Li, J.; Yang, J.; Zhu, Y.; Men, Y.; Zeng, Y.; Cai, Y.; Dong, C.; Dai, Z.; Zhang, X.; et al. Use of a promiscuous glycosyltransferase from Bacillus subtilis 168 for the enzymatic synthesis of novel Protopanaxatriol-type ginsenosides. J. Agric. Food Chem. 2018, 66, 943–949. [Google Scholar] [CrossRef]
- Dai, L.; Qin, L.; Hu, Y.; Huang, J.W.; Hu, Z.; Min, J.; Sun, Y.; Guo, R.T. Structural dissection of unnatural ginsenoside-biosynthetic UDP-glycosyltransferase Bs-YjiC from Bacillus subtilis for substrate promiscuity. Biochem. Biophys. Res. Commun. 2021, 534, 73–78. [Google Scholar] [CrossRef]
- Zhu, Y.; Li, J.; Peng, L.; Meng, L.; Diao, M.; Jiang, S.; Li, J.; **e, N. High-yield production of protopanaxadiol from sugarcane molasses by metabolically engineered Saccharomyces cerevisiae. Microb. Cell Fact. 2022, 21, 230. [Google Scholar] [CrossRef]
- Dai, Z.; Liu, Y.; Zhang, X.; Shi, M.; Wang, B.; Wang, D.; Huang, L.; Zhang, X. Metabolic engineering of Saccharomyces cerevisiae for production of ginsenosides. Metab. Eng. 2013, 20, 146–156. [Google Scholar] [CrossRef]
- Zhao, F.L.; Bai, P.; Nan, W.H.; Li, D.H.; Zhang, C.B.; Lu, C.Z.; Qi, H.S.; Lu, W. A modular engineering strategy for high-level production of protopanaxadiol from ethanol by Saccharomyces cerevisiae. AIChE J. 2018, 65, 866–874. [Google Scholar] [CrossRef]
- Choi, B.H.; Kang, H.J.; Kim, S.C.; Lee, P.C. Organelle engineering in yeast: Enhanced production of protopanaxadiol through manipulation of peroxisome proliferation in Saccharomyces cerevisiae. Microorganisms 2022, 10, 650. [Google Scholar] [CrossRef]
- Wang, Y.; Zhang, H.; Ri, H.C.; An, Z.; Wang, X.; Zhou, J.N.; Zheng, D.; Wu, H.; Wang, P.; Yang, J.; et al. Deletion and tandem duplications of biosynthetic genes drive the diversity of triterpenoids in Aralia elata. Nat. Commun. 2022, 13, 2224. [Google Scholar] [CrossRef] [PubMed]
- Zhuang, Y.; Yang, G.Y.; Chen, X.; Liu, Q.; Zhang, X.; Deng, Z.; Feng, Y. Biosynthesis of plant-derived ginsenoside Rh2 in yeast via repurposing a key promiscuous microbial enzyme. Metab. Eng. 2017, 42, 25–32. [Google Scholar] [CrossRef] [PubMed]
- Wu, Y.; Xu, S.; Gao, X.; Li, M.; Li, D.; Lu, W. Enhanced protopanaxadiol production from xylose by engineered Yarrowia lipolytica. Microb. Cell Fact. 2019, 18, 83. [Google Scholar] [CrossRef] [PubMed]
- Li, D.; Wu, Y.; Zhang, C.; Sun, J.; Zhou, Z.; Lu, W. Production of triterpene ginsenoside compound K in the non-conventional yeast Yarrowia lipolytica. J. Agric. Food. Chem. 2019, 67, 2581–2588. [Google Scholar] [CrossRef] [PubMed]
- Zhao, C.; Gao, X.; Liu, X.; Wang, Y.; Yang, S.; Wang, F.; Ren, Y. Enhancing biosynthesis of a ginsenoside precursor by self-assembly of two key enzymes in Pichia pastoris. J. Agric. Food Chem. 2016, 64, 3380–3385. [Google Scholar] [CrossRef]
- Sun, Z.; Meng, H.; Li, J.; Wang, J.; Li, Q.; Wang, Y.; Zhang, Z.S. Identification of novel knockout targets for improving terpenoids biosynthesis in Saccharomyces cerevisiae. PLoS ONE 2014, 9, e112615. [Google Scholar] [CrossRef]
- Dong, L.; Cheng, R.; **ao, L. Diversity and composition of bacterial endophytes among plant parts of Panax notoginseng. Chin. Med. 2018, 13, 41. [Google Scholar] [CrossRef]
- Fadiji, A.E.; Babalola, O.O. Metagenomics methods for the study of plant-associated microbial communities: A review. J. Microbiol. Methods 2020, 170, 105860. [Google Scholar] [CrossRef]
- Tang, K.; Zhang, Y.; Lin, D.; Han, Y.; Chen, C.A.; Wang, D.; Lin, Y.S.; Sun, J.; Zheng, Q.; Jiao, N. Cultivation-independent and cultivation-dependent analysis of microbes in the shallow-sea hydrothermal system off kueishantao island, Taiwan: Unmasking heterotrophic bacterial diversity and functional capacity. Front. Microbiol. 2018, 9, 279. [Google Scholar] [CrossRef]
- Misra, B.B.; Langefeld, C.D.; Olivier, M. Integrated omics: Tools, advances, and future approaches. J. Mol. Endocrinol. 2019, 62, R21–R45. [Google Scholar] [CrossRef]
- Hong, C.E.; Kim, J.U.; Lee, J.W.; Bang, K.H.; Jo, I.H. Metagenomic analysis of bacterial endophyte community structure and functions in Panax ginseng at different ages. 3 Biotech 2019, 9, 300. [Google Scholar] [CrossRef] [PubMed]
- Zhao, M.; Lin, Y.; Wang, Y.; Li, X.; Han, Y.; Wang, K.; Sun, C.; Wang, Y.; Zhang, M. Transcriptome analysis identifies strong candidate genes for ginsenoside biosynthesis and reveals its underlying molecular mechanism in Panax ginseng CA Meyer. Sci. Rep. 2019, 9, 615. [Google Scholar] [CrossRef] [PubMed]
- Jung, J.H.; Kim, H.Y.; Kim, H.S.; Jung, S.H. Transcriptome analysis of Panax ginseng response to high light stress. J. Ginseng Res. 2020, 44, 312–320. [Google Scholar] [CrossRef]
- Sun, W.; Qin, L.; Xue, H.; Yu, Y.; Ma, Y.; Wang, Y.; Li, C. Novel trends for producing plant triterpenoids in yeast. Crit. Rev. Biotechnol. 2019, 39, 618–632. [Google Scholar] [CrossRef] [PubMed]
- Liu, L.; Wei, Y.M.; Zhou, X.W.; Lin, J.; Sun, X.F.; Tang, K.X. Agrobacterium tumefaciens-mediated genetic transformation of the taxol-producing endophytic fungus Ozonium sp. EFY21. Genet. Mol. Res. 2013, 12, 2913–2922. [Google Scholar] [CrossRef]
- Noushahi, H.A.; Khan, A.H.; Noushahi, U.F.; Hussain, M.; Javed, T.; Zafar, M.; Batool, M.; Ahmed, U.; Liu, K.; Harrison, M.T.; et al. Biosynthetic pathways of triterpenoids and strategies to improve their biosynthetic efficiency. Plant Growth Regul. 2022, 97, 439–454. [Google Scholar] [CrossRef] [PubMed]
- Wang, P.; Wei, W.; Ye, W.; Li, X.; Zhao, W.; Yang, C.; Li, C.; Yan, X.; Zhou, Z. Synthesizing ginsenoside Rh2 in Saccharomyces cerevisiae cell factory at high-efficiency. Cell Discov. 2019, 5, 5. [Google Scholar] [CrossRef] [Green Version]
Structure | Name | R1 | R2 | R3 | R4 | Cellular Mechanisms | Ref. |
---|---|---|---|---|---|---|---|
Protopanaxadiol (PPD) Type | |||||||
Ra1 | Glc2-Glc | - | Glc6-Ara(p)4-Xyl | - | Not reported | ||
Rb1 | Glc2-Glc | - | Glc6-Glc | - | Inhibition of invasion and migration | [17,21] | |
Rb2 | Glc2-Glc | - | Glc6-Ara(p) | - | Inhibition of metastasis and proliferation | [17] | |
Rc | Glc2-Glc | - | Glc6-Ara(f) | - | Anti-proliferative activity | [19] | |
Rd | Glc2-Glc | - | Glc | - | Inhibit proliferation; Inhibit angiogenesis | [4] | |
Rg3 | Glc2-Glc | - | H | - | Repression of cell proliferation and induce apoptosis | [20,25] | |
Rh2 | Glc | - | H | - | Modulation of cell cycle; Regulation of inflammatory response molecules | [20,27] | |
F2 | Glc | - | Glc | - | Inhibit proliferation | [4] | |
CK | H | - | Glc | - | Modulation of growth factors and regulation of transcription factors; Induce apoptosis | [5,23] | |
Protopanaxatriol (PPT)-type | |||||||
Re | OH | Glc2-Rha | Glc | - | Not reported | ||
Rf | OH | Glc2-Glc | H | - | Cell cycle arrest and apoptosis | [19] | |
Rg1 | OH | Glc | Glc | - | Induce apoptosis; Repression of cell proliferation | [4,21] | |
Rg2 | OH | Glc2-Rha | H | - | Induce apoptosis; Repression of cell proliferation | [4] | |
Rh1 | OH | Glc | H | - | Regulation of gene coding for metalloproteinase; Repression of cell proliferation | [5,28] | |
F1 | OH | H | Glc | - | Modulation of death receptor | [29] | |
Notoginsenoside R1 | H | Glc2-Xyl Glc | Glc | - | Regulation of inflammatory response molecules | [19] | |
Ocotillol-type | |||||||
Majonoside R2 | OH | Glc2-Xyl | - | - | Not reported | ||
Vinaginsenoside R1 | OH | Ac-Glc2-Rha | - | - | Not reported | ||
Oleanolic acid type | |||||||
RO | GlcUA-Glc | - | - | Glc | Not reported | ||
ROA | GlcUA-Glc | - | - | Glc6-Glc | Not reported |
Host | Endophytic Strains | Type of Compounds/ Biotransformation Pathway | Major Media | Titer (mg/mL) | References |
Ginsenoside production by native endoyphtes | |||||
Aralia elata | Penicillium sp. | Rb2, Re | PDA liquid | 2.049 | [41] |
P. ginseng | Fusarium sp. | Total ginsenoside | PDA liquid | 0.181 | [40] |
Aspergillus sp. | 0.144 | ||||
Verticillium sp. | 0.144 | ||||
P. notoginseng | Fusarium sp. PN8 | Rb1, Rd, and Rg3 | PDA liquid | 1.061 | [39] |
Aspergillus sp. PN17 | Re, Rd, and Rg3 | 0.583 | |||
P. ginseng | Agrobacterium sp. | Rg3 | LL medium | 62.20 mg L−1 | [42] |
Rh2 | 18.60 mg L−1 | ||||
Biotransformation of major to rare ginsenosides by endophytes | |||||
P. ginseng | Arthrinium sp. | Rb1 → Rd → F2 → CK | PDA liquid | NA | [47] |
Burkholderia sp. | Rb1 → Rd → Rg3 | PDA liquid | NA | [48] | |
Flavobacterium sp. | Rb1 → Gyp-XVII | PDA liquid | NA | [49] | |
Platycodon grandiflorum | Luteibacter sp. | Rb1 → Rd → F2 | LL medium | 0.06692 | [50] |
Rb1 → Rd → F2 → CK | 0.03323 | ||||
Rb2 → CO → CY → CK | |||||
Rc →CMc1 → CMe → CK | |||||
Rg1 → Rh1 | NA | ||||
P. notoginseng | Fusarium oxysporum or Fusarium sp. | Rb1 → CK | LB medium | 0.02 | [38] |
Rb1 → F2 | 0.025 | ||||
Nodulisporium sp. | Re → 6-O-[α-L-Rhamnopyranosyl-(1→2)-β-D-glucopyranosyl]-20-O-β-glucopyranosyl-dammarane-3,6,12,20,24,25-hexaol | 0.125 | |||
Vinaginsenoside R13 | 0.09 | ||||
Fusarium oxysporum Nodulisporium sp. Bacillus sp. | Rg1 → Vinaginsenoside R22 | 0.065 | |||
Nodulisporium sp. | Rh1 → Pseudo-ginsenoside RT4 | 0.075 | |||
Fusarium oxysporum | Rh1 → PPT | 0.02 | |||
Brevundimonas sp. | Rh1 → Rg1 | 0.15 | |||
Rh1 → Vinaginsenoside R15 | 0.05 | ||||
Bacillus sp. | Rh1 → (20S)-3-O-β-D-glucopyranosyl-6-O-β-D-glucopyranosylprotopanaxatriol | 0.07 | |||
P. notoginseng | Enterobacter chengduensis | Rg1 → F1 | PDA medium | 13.24%; | [46] |
Trichoderma koningii | Rb1 → Rd | 40.00% | |||
Rb1 → Rg3 | 32.31%; | ||||
Penicillium chermesinum | Rb1 → Rd | 74.24% | |||
P. quinquefolius | Bacillus sp. G9y | Rc → Rd | Beef extract peptone | 100% | [51] |
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content. |
© 2023 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Chu, L.L.; Huy, N.Q.; Tung, N.H. Microorganisms for Ginsenosides Biosynthesis: Recent Progress, Challenges, and Perspectives. Molecules 2023, 28, 1437. https://doi.org/10.3390/molecules28031437
Chu LL, Huy NQ, Tung NH. Microorganisms for Ginsenosides Biosynthesis: Recent Progress, Challenges, and Perspectives. Molecules. 2023; 28(3):1437. https://doi.org/10.3390/molecules28031437
Chicago/Turabian StyleChu, Luan Luong, Nguyen Quang Huy, and Nguyen Huu Tung. 2023. "Microorganisms for Ginsenosides Biosynthesis: Recent Progress, Challenges, and Perspectives" Molecules 28, no. 3: 1437. https://doi.org/10.3390/molecules28031437