HDAC抑制剂对两株海洋真菌的代谢调控及其抗污损代谢产物的研究

Marine biofouling causes serious operational problems of maritime industries. There is an urgent need of non-toxic or less-toxic antifoulants for marine coating industry, which has important ecological significance and economic significance. Marine fungus is a good resource for obtaining non-toxic antifoulants. The application of epigenetic theory in filamentous fungi is becoming a powerful method to obtain novel natural products, and using small molecule inhibitors of epigenetic modification enzymes is one of the main methods. In the primary experiments, we have obtained new antifouling compounds from two antifouling marine fungal strains. In this project, we will continue to ① study the effects of two histone deacetylase inhibitors on the secondary metabolite biosynthesis of the two fungal strains, ②optimize the ferment conditions of the two fungal strains and ferment 100 L broths, respectively, ③quickly purify and identify antifouling compounds from their fermentation broths by combining chemical analysis with bioactivity tracking, ④study the structure-bioactivity relationships of new antifouling compounds, ⑤study the mode-of-action of new antifouling compounds, ⑥evaluate the field-antifouling activies of antifouling metabolites. The project will offer new natural non-toxic antifoulants and technical reserves for the development of marine new non-toxic antifouling paint, and offer implementation cases for the research methods of marine fungal metabolic regulation.

海洋生物污损已成为人们充分利用海洋资源所要迫切解决的首要问题之一。开发高效无毒的海洋新型防污剂需求紧迫，有重要生态意义和经济意义。海洋真菌是寻找高效无毒防污剂的重要源泉。表观遗传学正成为挖掘真菌次级代谢产物合成潜力的强有力手段，使用重要表观遗传修饰酶的小分子抑制剂是主要方法之一。申请人在前期工作中从两株有显著抗污损活性的海洋真菌（曲霉和青霉）中发现结构新颖的抗污损活性化合物。在此基础上，本项目拟开展：①研究两种组蛋白去乙酰化酶（HDAC）抑制剂对这两株真菌次级代谢生物合成的影响；②优化这两株活性菌株的发酵条件并进行100 L发酵；③快速分离鉴定这两株活性菌株发酵液中抗污损活性化合物；④研究抗污损先导化合物的构效关系；⑤探讨1种活性化合物的抗污损作用机制；⑥评价活性物质海上抗污损能力。本项目将为研制海洋新型无毒防污涂料提供高效无毒防污剂和技术储备，并为海洋真菌代谢调控研究方法提供实施案例。

海洋生物污损已成为人们充分利用海洋资源所要迫切解决的首要问题之一。开发高效无毒的海洋新型防污剂需求紧迫，有重要生态意义和经济意义。海洋真菌是寻找高效无毒防污剂的重要源泉。本项目深入开展了2株柳珊瑚共附生真菌活性菌株（即青霉Penicillium sp. SCSGAF0023、曲霉Aspergillus sp. SCSGAF 0076）中抗污损活性物质的研究，主要研究内容包括：① 探究了HDAC抑制剂辛二酰苯胺异羟肟酸和5-氮胞苷、甲醇、DMSO、以及多种氨基酸前体等对这两株菌发酵产物的影响。②选择土豆培养基分别采用静置和摇床方式对Penicillium sp. SCSGAF0023 进行了100 L的放大发酵，分别采用1#培养基和在1#培养基中添加0.05%甲醇和0.2% L-谷氨酸对Aspergillus sp. SCSGAF0076进行了100 L 发酵，对它们的发酵产物中化学成分进行了分离、鉴定、以及抗污损活性测试，探讨了个别类型活性化合物的抗污损和抗菌构型关系，并对曲霉SCSGAF 0076和细菌Bacillus sp. MNMCCE 001共培养发酵产物中抗菌和抗污损活性化合物进行了研究，从中共分离鉴定了114个化合物，包括33个新化合物，筛选到23个抗污损活性化合物。③ 阐释了一个大环内酰胺类化合物抗海洋解淀粉芽孢杆菌生物膜形成和一个蒽醌类化合物抗金黄色葡萄球菌生物膜形成的作用机制。④ 对分离自这两株菌的多个含量高的单体化合物和不同极性组分进行了野外海上抗污损测试，初价了它们的海上防污能力，有待改进涂料配方做进一步还上挂板防污测试。已发表27篇标注有本项目资助的文章（其中24篇SCI文章），申请1项国家发明专利，获授权3项国家发明专利，培养了博士和硕士研究生6名。为研制海洋新型无毒防污涂料提供了技术储备和活性化合物。