阿魏酸诱导污叉丝孔菌漆酶活性提升

Ferulic acid enhances the laccase activity of Dichomitus squalens

摘要:

目的 探究阿魏酸对污叉丝孔菌漆酶活性的影响及其分子机制，为微生物降解芳香族化合物提供理论依据。方法 通过在合成培养基中添加不同浓度的阿魏酸，观察其对污叉丝孔菌生长和漆酶活性的影响；利用转录组和蛋白组技术分析阿魏酸诱导下漆酶的转录和蛋白表达水平变化；通过RNAi技术敲降lcc3基因，检测其对漆酶活性及降解多种芳香族化合物能力的影响；利用自建的基于高斯萤光素酶和Nano萤光素酶的双萤光素酶系统鉴定lcc3基因的核心启动子区。结果 适量添加阿魏酸可显著提高污叉丝孔菌的漆酶活性。转录组和蛋白组分析发现，lcc3基因在阿魏酸诱导下转录水平和表达水平均显著上调。lcc3基因敲降菌株的漆酶活性及降解多种芳香族化合物的活性均显著下降，证实lcc3是污叉丝孔菌降解芳香族化合物相关的漆酶活性的关键基因。此外，通过双萤光素酶系统成功鉴定出lcc3基因的核心启动子区。结论 本研究揭示了阿魏酸对污叉丝孔菌漆酶活性的诱导作用及其分子机制，证明lcc3基因是阿魏酸诱导的漆酶活性及降解芳香族化合物的关键基因。鉴定lcc3基因核心启动子区为进一步地基因表达调控研究提供了基础，并为微生物漆酶降解芳香族化合物的应用提供了理论支持。

Abstract:

Objective To study the effect of ferulic acid on the laccase activity of Dichomitus squalens and its molecular mechanism, providing a theoretical basis for microbial degradation of aromatic compounds.Methods Ferulic acid was added to the synthetic medium at different concentrations to investigate its effects on the growth of D. squalens and laccase activity. Transcriptomic and proteomic analyses were performed to examine the changes in laccase transcription and protein expression levels induced by ferulic acid. The lcc3 gene was knocked down using RNAi technology, and the impact on laccase activity and the ability to degrade various aromatic compounds was assessed. A self-constructed dual-luciferase system based on Gaussia luciferase and Nano luciferase was employed to identify the core promoter region of the lcc3 gene.Results An appropriate concentration of ferulic acid can significantly enhance laccase activity in D. squalens. Transcriptomic and proteomic analyses revealed that the transcription and expression levels of the lcc3 gene were markedly up-regulated under ferulic acid induction. In the lcc3-gene-knocked-down strain, both laccase activity and the ability to degrade various aromatic compounds decreased significantly, confirming that lcc3 is a key gene for laccase-related activities in degrading aromatic compounds in D. squalens. Moreover, the dual-luciferase system successfully identified the core promoter region of the lcc3 gene.Conclusion This study first revealed that ferulic acid can induce D. squalens laccase activity and clarified the molecular mechanism. It was also proved that lcc3 is a key gene for ferulic acid-induced laccase activity and degradation of aromatic compounds in D. squalens. Identifying the core promoter region of lcc3 lays a foundation for gene expression regulation research, and these findings offer theoretical support for using microbial laccases to degrade aromatic compounds.