Skip to main content
SpringerLink
Log in

Efficient strategy for maintaining and enhancing the huperzine A production of Shiraia sp. Slf14 through inducer elicitation

  • Short Communication
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

Huperzine A (HupA), a naturally occurring lycopodium alkaloid, is a potent, highly specific and reversible inhibitor of acetylcholinesterase and is a potential treatment for Alzheimer’s disease. However, isolating HupA from Huperziaceae plants is inefficient; thus, extracting this compound from endophytic fungi may be more controllable and sustainable. However, the large-scale production of this chemical from endophytes is limited by the innate instability of endophytic fungi. In this study, we maintained the stability and viability of the HupA-producing endophytic fungus Shiraia sp. Slf14 and enhanced the HupA titers during fermentation by adding Huperzia serrata extracts (HSE), l-lysine, and acetic acid into the culture as inducers. Adding trace amounts of HupA clearly improved the HupA production of Shiraia sp. Slf14, reaching a maximum content of approximately 40 μg g−1. Moreover, the addition of HSE and l-lysine promoted HupA production in the flask fermentation. The aforementioned bioprocessing strategy may be potentially applied to other endophytic fungal culture systems for the efficient production of plant secondary metabolites.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

References

  1. Castillo M, Gupta RN, Ho YK, MacLean DB, Spenser ID (1970) Biosynthesis of lycopodine. Incorporation of Δ1-piperideine and of pelletierine. Can J Chem 48(18):2911–2918

    Article  CAS  Google Scholar 

  2. Castillo M, Gupta RN, MacLean DB, Spenser ID (1970) Biosynthesis of lycopodine from lysine and acetate. The pelletierine hypothesis. Can J Chem 48(12):1893–1903

    Article  CAS  Google Scholar 

  3. Comins DL, Al-awar RS (1995) Model studies toward the synthesis of the lycopodium alkaloid, phlegmarine. J Org Chem 60(3):711–716

    Article  CAS  Google Scholar 

  4. Hemscheidt T, Spenser ID (1993) Biosynthesis of lycopodine: incorporation of acetate via an intermediate with C2v symmetry. J Am Chem Soc 115(7):3020–3021

    Article  CAS  Google Scholar 

  5. Ju Z, Wang J, Pan SL (2009) Isolation and preliminary identification of the endophytic fungi which produce Huperzine A from four species in Huperziaceae and determination of Huperzine A by HPLC. Fudan Univ J Med Sci 36(4):445–449

    CAS  Google Scholar 

  6. Koshiba T, Yokoshima S, Fukuyama T (2009) Total synthesis of (−)-huperzine A. Org Lett 11(22):5354–5356

    Article  CAS  PubMed  Google Scholar 

  7. Kumar A, Patil D, Rajamohanan PR, Ahmad A (2013) Isolation, purification and characterization of vinblastine and vincristine from endophytic fungus Fusarium oxysporum isolated from Catharanthus roseus. PLoS One 8(9):e71805

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  8. Kusari S, Košuth J, Čellárová E, Spiteller M (2011) Survival-strategies of endophytic Fusarium solani against indigenous camptothecin biosynthesis. Fungal Ecol 4(3):219–223

    Article  Google Scholar 

  9. Li WK, Zhou JY, Lin ZW, Hu ZB (2007) Study on fermentation condition for production of huperzine A from endophytic fungus 2F09P03B of Huperzia serrata. Chin Med Biotechnol 2(4):254–259

    Google Scholar 

  10. Luo H, Li Y, Sun C, Wu Q, Song J, Sun Y, Steinmetz A, Chen S (2010) Comparison of 454-ESTs from Huperzia serrata and Phlegmariurus carinatus reveals putative genes involved in lycopodium alkaloid biosynthesis and developmental regulation. BMC Plant Biol 10(1):209–224

    Article  PubMed Central  PubMed  Google Scholar 

  11. Luo H, Sun C, Li Y, Wu Q, Song J, Wang D, Jia X, Li R, Chen S (2010) Analysis of expressed sequence tags from the Huperzia serrata leaf for gene discovery in the areas of secondary metabolite biosynthesis and development regulation. Physiol Plantarum 139(1):1–12

    Article  CAS  Google Scholar 

  12. Ma X, Gang DR (2004) The lycopodium alkaloids. Nat Prod Rep 21(6):752–772

    Article  CAS  PubMed  Google Scholar 

  13. Ma X, Gang DR (2008) In vitro production of huperzine A, a promising drug candidate for Alzheimer’s disease. Phytochemistry 69(10):2022–2028

    Article  CAS  PubMed  Google Scholar 

  14. Ma X, Tan C, Zhu D, Gang DR, Xiao P (2007) Huperzine A from Huperzia species—an ethnopharmacolgical review. J Ethnopharmacol 113(1):15–34

    Article  CAS  PubMed  Google Scholar 

  15. Marshall WD, Nguyen TT, MacLean DB, Spenser ID (1975) Biosynthesis of lycopodine. The question of the intermediacy of piperidine-2-acetic acid. Can J Chem 53(1):41–50

    Article  CAS  Google Scholar 

  16. Nyembo L, Goffin A, Hootele C, Braekman J-C (1978) Phlegmarine, a likely key intermediate in the biosynthesis of the Lycopodium alkaloids. Can J Chem 56(6):851–856

    Article  CAS  Google Scholar 

  17. Qian L, Ji R (1989) A total synthesis of (±)-huperzine A. Tetrahedron Lett 30(16):2089–2090

    Article  CAS  Google Scholar 

  18. Rafii MS, Walsh S, Little JT, Behan K, Reynolds B, Ward C, Jin S, Thomas R, Aisen PS (2011) A phase II trial of huperzine A in mild to moderate Alzheimer disease. Neurology 76(16):1389–1394

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  19. Stierle A, Strobel G, Stierle D (1993) Taxol and taxane production by Taxomyces andreanae, an endophytic fungus of Pacific yew. Science 260(5105):214–216

    Article  CAS  PubMed  Google Scholar 

  20. Sudhakar T, Dash SK, Rao RR, Srinivasan R, Zacharia S, Atmanand MA, Subramaniam BR, Nayak S (2013) Do endophytic fungi possess pathway genes for plant secondary metabolites? Curr Sci India 104(2):178–182

    Google Scholar 

  21. Szypuła W, Pietrosiuk A, Suchocki P, Olszowska O, Furmanowa M, Kazimierska O (2005) Somatic embryogenesis and in vitro culture of Huperzia selago shoots as a potential source of huperzine A. Plant Sci 168(6):1443–1452

    Article  Google Scholar 

  22. Wang Y, Zeng QG, Zhang ZB, Yan RM, Wang LY, Zhu D (2011) Isolation and characterization of endophytic huperzine A-producing fungi from Huperzia serrata. J Ind Microbiol Biot 38(9):1267–1278. doi:10.1007/s10295-010-0905-4

    Article  CAS  Google Scholar 

  23. Yang X, Guo S, Zhang L, Shao H (2003) Select of producing podophyllotoxin endophytic fungi from podophyllin plant. Nat Product Res Dev 15(5):419–422

    CAS  Google Scholar 

  24. Yi M-H, Kim J-A, Kim J-M, Park J-A, Kim B-T, Park S-M, Yang M-S, Hwang K-J, Kim D-H (2011) Characterization of a mutant strain of a filamentous fungus Cladosporium phlei for the mass production of the secondary metabolite phleichrome. J Microbiol 49(4):680–683

    Article  CAS  PubMed  Google Scholar 

  25. Zhang ZB, Zeng QG, Yan RM, Wang Y, Zou ZR, Zhu D (2011) Endophytic fungus Cladosporium cladosporioides LF70 from Huperzia serrata produces huperzine A. World J Microbiol Biotechnol 27(3):479–486

    Article  CAS  Google Scholar 

  26. Zhao X-M, Wang Z-Q, Shu S-H, Wang W-J, Xu H-J, Ahn Y-J, Wang M, Hu X (2013) Ethanol and methanol can improve huperzine A production from endophytic Colletotrichum gloeosporioides ES026. PLoS One 8(4):e61777

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  27. Zhu D, Wang J, Zeng Q, Zhang Z, Yan R (2010) A novel endophytic huperzine A-producing fungus, Shiraia sp. Slf14, isolated from Huperzia serrata. J Appl Microbiol 109(4):1469–1478

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by the National Natural Science Foundation of China (Grant Nos. 31300051, 21066014 and 81260617), China Postdoctoral Science Foundation (2012M511454), Jiangxi Natural Science Foundation of China (20114BAB204020), and Major Science and Technology Subject of Jiangxi Province (2010AZD00307). We appreciate the help of Prof. Lehua Zhang from the Lushan Botanical Garden of China for providing the plant materials. We are also grateful for the constructive comments of anonymous reviewers.

Author information

Authors and Affiliations

  1. Key Laboratory of Protection and Utilization of Subtropic Plant Resources of Jiangxi Province, Jiangxi Normal University, Nanchang, 330022, China

    Riming Yan, Zhibin Zhang, Huilin Yang, Qinggui Zeng & Du Zhu

  2. Key Laboratory for Research on Active Ingredients in Natural Medicine of Jiangxi Province, Yichun University, Yichun, 336000, China

    Riming Yan

  3. Jiangxi Science and Technology Normal University, Nanchang, 330013, China

    Ya Wang & Du Zhu

Authors
  1. Riming Yan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Zhibin Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ya Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Huilin Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Qinggui Zeng
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Du Zhu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Du Zhu.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Yan, R., Zhang, Z., Wang, Y. et al. Efficient strategy for maintaining and enhancing the huperzine A production of Shiraia sp. Slf14 through inducer elicitation. J Ind Microbiol Biotechnol 41, 1175–1179 (2014). https://doi.org/10.1007/s10295-014-1461-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10295-014-1461-0

Keywords

Search

Navigation