Vats Sheetal, Saxena Sanjai
Department of Biotechnology, Thapar Institute of Engineering and Technology, Patiala, Punjab, 147004, India.
Arch Microbiol. 2023 May 8;205(6):224. doi: 10.1007/s00203-023-03575-z.
Aggregation of α-synuclein into toxic oligomeric structures has been implicated in the pathogenesis of Parkinson's disease via several key stages of fibrillation, oligomerization, and aggregation. Disaggregation or prevention of aggregation has garnered a lot of attention as a therapeutic strategy to prevent or delay the progression of Parkinson's disease. It has been recently established that certain polyphenolic compounds and catechins present in plants and tea extracts exhibit the potential to inhibit the α-synuclein aggregation. However, their copious supply for therapeutic development is still unsolved. Herein, we report for the first time the disaggregation potential of α-synuclein by an endophytic fungus residing in tea leaves (Camellia sinensis). Briefly, a recombinant yeast expressing α-synuclein was used for pre-screening of 53 endophytic fungi isolated from tea using anti-oxidant activity as a marker for the disaggregation of the protein. One isolate #59CSLEAS exhibited 92.4% reduction in production of the superoxide ions, which were similar to the already established α-synuclein disaggregator, Piceatannol exhibiting 92.8% reduction. Thioflavin T assay further established that #59CSLEAS decreased the oligomerization of α-synuclein by 1.63-fold. Subsequently Dichloro-dihydro-fluorescein diacetate-based fluorescence assay exhibited a reduction in total oxidative stress in the recombinant yeast in the presence of fungal extract, thereby indicating the prevention of oligomerization. Oligomer disaggregation potential of the selected fungal extract was found to be 56.5% as assessed by sandwich ELISA assay. Using morphological as well as molecular methods, the endophytic isolate #59CSLEAS was identified as Fusarium sp. The sequence was submitted in the Genbank with accession number ON226971.1.
α-突触核蛋白聚集成有毒的寡聚体结构，已通过纤维化、寡聚化和聚集的几个关键阶段，与帕金森病的发病机制相关。作为预防或延缓帕金森病进展的治疗策略，解聚或防止聚集已引起广泛关注。最近已证实，植物和茶叶提取物中存在的某些多酚化合物和儿茶素具有抑制α-突触核蛋白聚集的潜力。然而，它们用于治疗开发的大量供应问题仍未解决。在此，我们首次报道了一种存在于茶叶（茶树）中的内生真菌对α-突触核蛋白的解聚潜力。简要地说，使用表达α-突触核蛋白的重组酵母，以抗氧化活性作为蛋白质解聚的标志物，对从茶叶中分离的53种内生真菌进行预筛选。一种分离株#59CSLEAS使超氧离子的产生减少了92.4%，这与已确立的α-突触核蛋白解聚剂白藜芦醇相似，后者使超氧离子的产生减少了92.8%。硫黄素T分析进一步证实，#59CSLEAS使α-突触核蛋白的寡聚化降低了1.63倍。随后，基于二氯二氢荧光素二乙酸酯的荧光分析表明，在存在真菌提取物的情况下，重组酵母中的总氧化应激降低，从而表明防止了寡聚化。通过夹心ELISA分析评估，所选真菌提取物的寡聚体解聚潜力为56.5%。使用形态学和分子方法，将内生分离株#59CSLEAS鉴定为镰刀菌属。该序列已提交至Genbank，登录号为ON226971.1。
