| 窦敏,刘婕,范爱丽,等.海洋真菌A.sclerotiorum LZDX 33-4中吲哚二酮哌嗪类生物碱基因簇异源表达研究[J].中国海洋药物,2025,44(1):65-70. |
| 海洋真菌A.sclerotiorum LZDX 33-4中吲哚二酮哌嗪类生物碱基因簇异源表达研究 |
| Heterologous expression of notoamide-type alkaloids’ gene cluster from marine fungus Aspergillus sclerotiorum LZDX-33 |
| 投稿时间：2023-09-21 修订日期：2023-12-01 |
| DOI：10.13400/j.cnki.cjmd.2025.01.008 |
| 中文关键词: 海洋真菌 Aspergillus sclerotiorum 二酮哌嗪类生物碱 异源表达 |
| English Keywords:marine fungi Aspergillus sclerotiorum diketopiperazine alkaloid heterologous expression |
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| 中文摘要: |
| 目的 运用分子生物学的方法，对深海来源真菌Aspergillus sclerotiorum LZDX 33-4中吲哚二酮哌嗪类生物碱的产率提高进行研究。方法 利用生物信息学方法从A. sclerotiorum LZDX 33-4中钓取到负责吲哚二酮哌嗪类生物碱合成的基因簇stm；利用异源表达策略在构巢曲霉中构建全簇异源表达菌株；利用十八烷基硅烷键合硅胶（ODS）和半制备高效液相色谱（HPLC）等色谱学方法对异源表达菌株提取物中吲哚二酮哌嗪类生物碱进行靶向分离纯化；利用UV、NMR等波谱学技术手段，结合文献数据比对，确定化合物的结构。结果 从异源表达菌株中分离鉴定了5个化合物，分别为(+)-versicolamide B(1)、taichunamide E(2)、notoamide B(3)、taichunamide A(4)、notoamide D(5)。其中化合物1-5的产率分别为7.27mg/L，1.87mg/L，0.33mg/L，2.53mg/L和2.03mg/L。结论 通过构建异源表达菌株，分离鉴定了5个吲哚二酮哌嗪生物碱类化合物。与野生型菌株相比，极大的提高了该类化合物的产率。 |
| English Summary: |
| Objective The heterologous production of notoamide-type alkaloids from Aspergillus sclerotiorum LZDX 33-4 was studied by molecular biological methods. Methods The gene cluster stm responsible for notoamides’ biosynthesis was found in the genome of A. sclerotiorum LZDX 33-4 by bioinformatics analysis. The heterologous expression strategy was used to construct the whole cluster of expression strain in A. nidulatus. The extracts of heterologously expressed strains were isolated and purified by ODS and semi-preparative high performance liquid chromatography (HPLC). The structure of the compound was determined by UV, NMR and other spectroscopic techniques combined with the comparison of literature data. Results Five compounds were isolated and identified from the mutant strain, including (+)-versicolamide B(1)、taichunamide E(2)、notoamide B(3)、taichunamide A(4)、notoamide D(5). The yields of 1-5 were 7.27mg/L, 1.87mg/L, 0.33mg/L, 2.53mg/L and 2.03mg/L, respectively. Conclusion Overexpressed strain was constructed by heterologous expression methods, and 5 compounds were isolated and identified. Compared with the wild-type strain, and the production efficiency of notoamide-type alkaloids was greatly increased. |
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