Abstract
Highly prized huperzine A (Hup A), a natural alkaloid formerly isolated from the Chinese medicinal plant Huperzia serrata, has been widely used for the treatment of Alzheimer disease, inspiring us to search for endophytic fungi that produce this compound. In this study, we obtained the C17 fungus isolate from the Mexican club moss Phlegmariurus taxifolius, which produced a yield of 3.2 μg/g Hup A in mycelial dry weight, when cultured in potato dextrose broth medium. The C17 isolate was identified as belonging to the genus Fusarium with reference to the colony´s morphological characteristics and the presence of macroconidia and microconidia structures; and this was confirmed by DNA-barcoding analysis, by amplifying and sequencing the ribosomal internal transcribed spacer (rITS).
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References
Patterson C (2018) World Alzheimer Report 2018—The state of the art of dementia research: New frontiers. Alzheimer’s Disease International, London. https://www.alz.co.uk/research/world-report-2018. Accessed 30 June 2019
Bai DL, Tang XC, He XC (2000) Huperzine A, a potential therapeutic agent for treatment of Alzheimer’s disease. Curr Med Chem 7(3):355–374. https://doi.org/10.2174/0929867003375281
Darvesh S, Walsh R, Kumar R, Caines A, Roberts S, Magee D, Rockwood K, Martin E (2003) Inhibition of human cholinesterases by drugs used to treat Alzheimer disease. Alzheimer Dis Assoc Disord 17(2):117–126. https://doi.org/10.1097/00002093-200304000-00011
Wang R, Yan H, Tang XC (2006) Progress in studies of huperzine A, a natural cholinesterase inhibitor from Chinese herbal medicine. Acta Pharmacol Sin 27(1):1–26. https://doi.org/10.1111/j.1745-7254.2006.00255.x
Testo W, Øllgaard B, Field A, Almeida T, Kessler M, Barrington D (2018) Phylogenetic systematics, morphological evolution, and natural groups in neotropical Phlegmariurus (Lycopodiaceae). Mol Phylogenet Evol 125:1–13. https://doi.org/10.1016/j.ympev.2018.03.016
Ma X, Tan C, Zhu D, Gang DR, Xiao P (2007) Huperzine A from Huperzia species—an ethnopharmacological review. J Ethnopharmacol 113(1):15–34. https://doi.org/10.1016/j.jep.2007.05.030
Ma X, Tan C, Zhu D, Gang DR (2006) A survey of potential huperzine A natural resources in China: the Huperziaceae. J Ethnopharmacol 104(1–2):54–67. https://doi.org/10.1016/j.jep.2005.08.042
Ma X, Gang DR (2004) The Lycopodium alkaloids. Nat Prod Rep 21(6):752–772. https://doi.org/10.1039/b409720n
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. https://doi.org/10.1126/science.8097061
Strobel G, Daisy B (2003) Bioprospecting for microbial endophytes and their natural products. Microbiol Mol Bio Rev 67(4):491–502. https://doi.org/10.1128/MMBR.67.4.491-502.2003
Gouda S, Das G, Sen SK, Shin HS, Patra JK (2016) Endophytes: a treasure house of bioactive compounds of medicinal importance. Front Microbiol 7:1538. https://doi.org/10.3389/fmicb.2016.01538
Zhao J, Zhou L, Wang J, Shan T, Zhong L, Liu X, Gao X (2010) Endophytic fungi for producing bioactive compounds originally from their host plants. Curr Res Technol Educ Top Appl Microbiol Microbial Biotechnol 1:567–576
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. https://doi.org/10.1111/j.1365-2672.2010.04777.x
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. https://doi.org/10.1007/s11274-010-0476-6
Shu S, Zhao X, Wang W, Zhang G, Cosoveanu A, Ahn Y, Wang M (2014) Identification of a novel endophytic fungus from Huperzia serrata which produces huperzine A. World J Microbiol Biotechnol 30(12):3101–3109. https://doi.org/10.1007/s11274-014-1737-6
Zhang FF, Wang MZ, Zheng YX, Liu HY, Zhang XQ, Wu SS (2015) Isolation and characterization of endophytic huperzine A-producing fungi from Phlegmariurus phlegmaria. Microbiology 84(5):701–709. https://doi.org/10.1134/S0026261715050185
Bayman P, Lebrón LL, Tremblay RL, Lodge DJ (1997) Variation in endophytic fungi from roots and leaves of Lepanthes (Orchidaceae). New Phytol 135:143–149
Leslie JF, Summerell BA, Bullock S (2006) The Fusarium laboratory manual. Blackwell-Wiley, USA
Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22(22):4673–4680. https://doi.org/10.1093/nar/22.22.4673
Kumar S, Stecher G, Tamura K (2016) MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol 33(7):1870–1874. https://doi.org/10.1093/molbev/msw054
Ellman GL, Courtney KD, Andres JV, Featherstone RM (1961) A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol 7:88–95. https://doi.org/10.1016/0006-2952(61)90145-9
Eberl F, Uhe C, Unsicker SB (2019) Friend or foe? The role of leaf-inhabiting fungal pathogens and endophytes in tree-insect interactions. Fungal Ecol 38:104–112. https://doi.org/10.1016/j.funeco.2018.04.003
Domsch KH, Gams W, Anderson TH (1980) “Fusarium”. Compendium of soil fungi. Academic Press, London, pp 333–337
Tsuchinari M, Shimanuki K, Hiramatsu F, Murayama T, Koseki T, Shiono Y (2007) Fusapyridons A and B, novel pyridone alkaloids from an endophytic fungus, Fusarium sp. YG-45. Z Naturforsch B 62(9):1203–1207. https://doi.org/10.1515/znb-2007-0916
Kumara PM, Zuehlke S, Priti V, Ramesha BT, Shweta S, Ravikanth G, Vasudeva R, Santhoshkumar TR, Spiteller M, Shaanker RU (2012) Fusarium proliferatum, an endophytic fungus from Dysoxylum binectariferum Hook.f, produces rohitukine, a chromane alkaloid possessing anti-cancer activity. Antonie Van Leeuwenhoek 101(2):323–329. https://doi.org/10.1007/s10482-011-9638-2
Wen H, Li Y, Liu X, Ye W, Yao X, Che Y (2015) Fusagerins A-F, new alkaloids from the fungus Fusarium sp. Nat Prod Bioprospect 5(4):195–203. https://doi.org/10.1007/s13659-015-0067-1
Nenkep V, Yun K, Son BW (2016) Oxysporizoline, an antibacterial polycyclic quinazoline alkaloid from the marine-mudflat-derived fungus Fusarium oxysporum. J Antibiot 69(9):709–711. https://doi.org/10.1038/ja.2015.137
Zhao XM, Wang ZQ, Shu SH, Wang WJ, Xu HJ, Ahn YJ, Wang M, Hu X (2013) Ethanol and methanol can improve huperzine A production from endophytic Colletotrichum gloeosporioides ES026. PLoS ONE 8(4):e61777. https://doi.org/10.1371/journal.pone.0061777
Ma X, Tan C, Zhu D, Gang DR (2005) Is there a better source of huperzine A than Huperzia serrata? Huperzine A content of Huperziaceae species in China. J Agric Food Chem 53(5):1393–1398. https://doi.org/10.1021/jf048193n
Dong LH, Fan SW, Ling QZ, Huang BB, Wei ZJ (2014) Identification of huperzine A-producing endophytic fungi isolated from Huperzia serrata. World J Microbiol Biotechnol 30(3):1011–1017. https://doi.org/10.1007/s11274-013-1519-6
Su J, Yang M (2015) Huperzine A production by Paecilomyces tenuis YS-13, an endophytic fungus isolated from Huperzia serrata. Nat Prod Res 29(11):1035–1041. https://doi.org/10.1080/14786419.2014.980245
Zaki AG, El-Shatoury EH, Ahmed AS, Al-Hagar OEA (2019) Production and enhancement of the acetylcholinesterase inhibitor, huperzine A, from an endophytic Alternaria brassicae AGF041. Appl Microbiol Biot 103:5867–5878. https://doi.org/10.1007/s00253-019-09897-7
Zan JU, Wang J, Pan S (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 36:445–449
Acknowledgements
This research was supported by CONACyT (Nos. 156276 and 222714) developed at the Centro de Investigación en Biotecnología of the Universidad Autónoma del Estado de Morelos, Mexico. Thanks are due to Ariadna Zenil for support in the HPLC analysis. OLCM acknowledges the receipt of a Ph.D. fellowship No. 230949 from CONACYT.
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Cruz-Miranda, O., Folch-Mallol, J., Martínez-Morales, F. et al. Identification of a Huperzine A-producing endophytic fungus from Phlegmariurus taxifolius. Mol Biol Rep 47, 489–495 (2020). https://doi.org/10.1007/s11033-019-05155-1
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DOI: https://doi.org/10.1007/s11033-019-05155-1