Huperzine A (Hup A) is an important drug for treating Alzheimer's disease (AD) and mainly extracted from the Huperzia serrata (Thunb.) Trevis. (Lycopodiaceae) (HS). Nevertheless, the content of Hup A in HS is very low of 0.007% with growing circle of 8 to 10 years, and the chemical synthesis of Hup A still has some insurmountable limitations in the industrialized production. So, the available resources of Hup A for clinical treatment of AD are scarce. The purpose of this work was to construct a biosynthesis platform based on the endophytic fungi from HS. In this work, five endophytic fungi Mucor racemosus NSH-D, Mucor fragilis NSY-1, Fusarium verticillioides NSH-5, Fusarium oxysporum NSG-1, and Trichoderma harzianum NSW-V were firstly found and isolated from the Chinese folk medicine HS, which were identified according to their morphological characteristics and nuclear ribosomal DNA ITS sequences. The highest efficient fungus could effectively biosynthesize Hup A in a liquid culture of 319.8 ± 0.17 mg/L which were 112 times higher than that of other reported conventional endophytic fungi. Moreover, these fungi with higher hereditary stability could possess the initial expressing ability of Hup A after 40 generations, and the expressed Hup A from these biosynthesis systems has prior physicochemical properties, a better inhibition activity of acetylcholinesterase and a lower cytotoxicity compared with the listed active pharmaceutical ingredients (APIs) of Hup A. These results provide promising alternative resources for producing Hup A at an industrial scale by biosynthesis, and it may also shed light on millions of AD patients. KEY POINTS: • Five novel endophytic fungi with high stability could highly express prior Hup A Graphical abstract.

中文翻译：

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从表达石杉碱甲的石杉石杉分离的五种新型高效内生真菌用于治疗阿尔茨海默氏病。

石杉碱甲（Hup A）是一种用于治疗阿尔茨海默氏病（AD）的重要药物，主要从锯齿石杉（Thunb。）Trevis中提取。（茄科）（HS）。但是，HS中Hup A的含量很低，仅为0.007％，生长周期为8至10年，Hup A的化学合成在工业化生产中仍然存在一些不可克服的局限性。因此，Hup A可用于AD临床治疗的可用资源很少。这项工作的目的是构建基于HS的内生真菌的生物合成平台。在这项工作中，首先从中药HS中发现并分离了5种内生真菌消旋毛霉NSH-D，脆弱类毛霉NSY-1，轮状镰刀菌NSH-5，尖孢镰刀NSG-1和哈茨木霉NSW-V，根据它们的形态特征和核糖体DNA ITS序列鉴定。最高效率的真菌可以在319.8±0.17 mg / L的液体培养物中有效地生物合成Hup A，是其他报道的常规内生真菌的112倍。而且，具有较高遗传稳定性的这些真菌可以在40代后具有Hup A的初始表达能力，并且与所列活性物质相比，从这些生物合成系统表达的Hup A具有先验的理化特性，更好的乙酰胆碱酯酶抑制活性和较低的细胞毒性。 Hup A的药物成分（API）。这些结果为通过生物合成在工业规模生产Hup A提供了有前途的替代资源，它也可能为数百万的AD患者提供了启示。