State Key Laboratory of Applied Microbiology Southern China, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Institute of Microbiology, Guangdong Academy of Sciences, China.
State Key Laboratory of Applied Microbiology Southern China, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Institute of Microbiology, Guangdong Academy of Sciences, China.
Int J Biol Macromol. 2024 Nov;280(Pt 2):135746. doi: 10.1016/j.ijbiomac.2024.135746. Epub 2024 Sep 16.
Type D trichothecene toxins represent a class of macrocyclic trichothecene toxins with significant cytotoxicities towards human and crops. These toxins can also be used as anti-tumor compounds by the combination of antibody-drug conjugate. Therefore, it is urgent to investigate the biosynthetic routine of type D trichothecene toxins and explore type D trichothecene toxin-resistant genes, in order to ameliorate the hazard of trichothecene toxins and to facilitate the heterologous expression of toxin-biosynthetic cluster. In this study, tri18 gene was firstly knocked out in Paramyrothecium roridum, leading to the complete absence of type D trichothecene toxin epiroridin E, which can be restored by the complement of tri18 gene. Additionally, the knockout of tri18 gene led to a significant reduction in the pathogenicity of P. roridum towards pumpkin. Meanwhile, the enzymatic properties of Tri18 protein towards trichothecene deoxynivalenol (DON) toxin were also characterized. Moreover, tri3 and tri17KR with broad spectrum toxin-resistance function within the tri cluster were initially discovered through heterologous expression in toxin-sensitive Saccharomyces cerevisiae. And this study provides innovative type D trichothecene toxin resistant enzymes, which can provides green platform for the production of type D trichothecene toxins, thus promoting the application of these toxins in biomedical field.
D 型单端孢霉烯族毒素属于一类具有显著细胞毒性的大环单端孢霉烯族毒素，对人和作物都有很大的毒性。这些毒素也可以通过抗体药物偶联物的结合作为抗肿瘤化合物使用。因此，迫切需要研究 D 型单端孢霉烯族毒素的生物合成途径，并探索 D 型单端孢霉烯族毒素抗性基因，以减轻单端孢霉烯族毒素的危害，并促进毒素生物合成基因簇的异源表达。在本研究中，首先敲除了疣孢漆斑菌中的 tri18 基因，导致 D 型单端孢霉烯族毒素 epiroridin E 的完全缺失，而 tri18 基因的互补可以恢复其产生。此外，tri18 基因的敲除导致疣孢漆斑菌对南瓜的致病性显著降低。同时，还对 Tri18 蛋白对单端孢霉烯族脱氧雪腐镰刀菌烯醇（DON）毒素的酶学特性进行了研究。此外，还通过在敏感的酿酒酵母中异源表达，初步发现了 tri3 和 tri17KR 这两个具有广谱毒素抗性功能的 tri 基因簇内的基因。本研究提供了创新的 D 型单端孢霉烯族毒素抗性酶，为 D 型单端孢霉烯族毒素的生产提供了绿色平台，从而促进了这些毒素在生物医学领域的应用。
