Laboratory of Environmental Biotechnology, Institute of Microbiology ASCR,v.v.i., Videnska 1083, 14220 Prague, Czech Republic.
Microb Biotechnol. 2013 May;6(3):300-6. doi: 10.1111/1751-7915.12007. Epub 2012 Nov 22.
Fungal, ligninolytic enzymes have attracted a great attention for their bioremediation capabilities. A deficient knowledge of regulation of enzyme production, however, hinders the use of ligninolytic fungi in bioremediation applications. In this work, a transcriptional analyses of laccase and manganese peroxidase (MnP) production by two white rots was combined with determination of pI of the enzymes and the evaluation of 17α-ethinyloestradiol (EE2) degradation to study regulation mechanisms used by fungi during EE2 degradation. In the cultures of Trametes versicolor the addition of EE2 caused an increase in laccase activity with a maximum of 34.2 ± 6.7 U g⁻¹ of dry mycelia that was observed after 2 days of cultivation. It corresponded to a 4.9 times higher transcription levels of a laccase-encoding gene (lacB) that were detected in the cultures at the same time. Simultaneously, pI values of the fungal laccases were altered in response to the EE2 treatment. Like T. versicolor, Irpex lacteus was also able to remove 10 mg l⁻¹ EE2 within 3 days of cultivation. While an increase to I. lacteus MnP activity and MnP gene transcription levels was observed at the later phase of the cultivation. It suggests another metabolic role of MnP but EE2 degradation.
真菌木质素降解酶因其生物修复能力而受到极大关注。然而,由于对酶生产调控机制的了解不足,限制了木质素降解真菌在生物修复应用中的使用。在这项工作中,我们结合了两种白腐真菌漆酶和锰过氧化物酶(MnP)生产的转录分析,以及酶等电点(pI)的测定和 17α-乙炔基雌二醇(EE2)降解的评估,研究真菌在 EE2 降解过程中使用的调控机制。在糙皮侧耳的培养物中,添加 EE2 导致漆酶活性增加,最大增加量为 2 天培养后干菌丝的 34.2±6.7 U g⁻¹。这与同时检测到的编码基因(lacB)转录水平增加了 4.9 倍相对应。同时,真菌漆酶的 pI 值也因 EE2 处理而发生变化。与糙皮侧耳相似,乳白栓菌也能够在 3 天的培养时间内去除 10mg l⁻¹ 的 EE2。虽然在培养后期观察到 I. lacteus 的 MnP 活性和 MnP 基因转录水平增加,但这表明 MnP 具有另一种代谢作用,但不是 EE2 降解。