Yang Lu, Wu Guangwei, Meng Fanyue, Ran Huomiao, Yin Wenbing, Li Wei, Liu Xiaoqing
College of Life Science, Capital Normal University, Beijing 100048, China.
Jiangsu Co-Innovation Center of Efficient Processing and Utilization of Forest Resources, Jiangsu Key Lab of Biomass-Based Green Fuels and Chemicals, College of Chemical Engineering, Nanjing Forestry University, Nanjing 210037, China.
Int J Mol Sci. 2022 Mar 28;23(7):3686. doi: 10.3390/ijms23073686.
The strategies of genetic dereplication and manipulation of epigenetic regulators to activate the cryptic gene clusters are effective to discover natural products with novel structure in filamentous fungi. In this study, a combination of genetic dereplication (deletion of pesthetic acid biosynthetic gene, ) and manipulation of epigenetic regulators (deletion of histone methyltransferase gene and histone deacetylase gene ) was developed in plant endophytic fungus . The deletion of with and/or led to isolation of 1 novel compound, pestaloficiol X (), as well as another 11 known compounds with obvious yield changes. The proposed biosynthesis pathway of pestaloficiol X was speculated using comparative analysis of homologous biosynthetic gene clusters. Moreover, phenotypic effects on the conidial development and response to oxidative stressors in the mutants were explored. Our results revealed that the new strain with deletion of or in Δ background host can neutralise the hyperformation of conidia in the mutant, and that the Δ Δ mutant was generally not sensitive to oxidative stressors as much as the Δ Δ mutant in comparison with the single mutant Δ or the parental strains. This combinatorial approach can be applied to discover new natural products in filamentous fungi.
通过基因去重复和操纵表观遗传调控因子来激活隐秘基因簇的策略，对于在丝状真菌中发现具有新结构的天然产物是有效的。在本研究中，在植物内生真菌中开发了一种基因去重复（删除佩斯托酸生物合成基因）和表观遗传调控因子操纵（删除组蛋白甲基转移酶基因和组蛋白脱乙酰酶基因）相结合的方法。删除与和/或导致分离出1种新化合物，即佩斯托菲醇X（），以及另外11种产量有明显变化的已知化合物。利用同源生物合成基因簇的比较分析推测了佩斯托菲醇X的拟生物合成途径。此外，还探索了突变体对分生孢子发育和氧化应激反应的表型效应。我们的结果表明，在Δ背景宿主中缺失或的新菌株可以中和突变体中分生孢子的过度形成，并且与单突变体Δ或亲本菌株相比，ΔΔ突变体通常对氧化应激不如ΔΔ突变体敏感。这种组合方法可用于在丝状真菌中发现新的天然产物。
