Torres-Farradá Giselle, Manzano León Ana M, Rineau François, Ledo Alonso Lucía L, Sánchez-López María I, Thijs Sofie, Colpaert Jan, Ramos-Leal Miguel, Guerra Gilda, Vangronsveld Jaco
Laboratory of Biotechnology, Department of Microbiology and Virology, Faculty of Biology, University of HavanaHavana, Cuba.
Department of Phytopathology, Research Institute for Tropical Fruit TreesHavana, Cuba.
Front Microbiol. 2017 May 23;8:898. doi: 10.3389/fmicb.2017.00898. eCollection 2017.
White-rot fungi (WRF) and their ligninolytic enzymes (laccases and peroxidases) are considered promising biotechnological tools to remove lignin related Persistent Organic Pollutants from industrial wastewaters and contaminated ecosystems. A high diversity of the genus has been reported in Cuba; in spite of this, the diversity of ligninolytic enzymes and their genes remained unexplored. In this study, 13 native WRF strains were isolated from decayed wood in urban ecosystems in Havana (Cuba). All strains were identified as sp. using a multiplex polymerase chain reaction (PCR)-method based on ITS sequences. All sp. strains produced laccase enzymes at higher levels than non-specific peroxidases. Native-PAGE of extracellular enzymatic extracts revealed a high diversity of laccase isozymes patterns between the strains, suggesting the presence of different amino acid sequences in the laccase enzymes produced by these strains. We determined the diversity of genes encoding laccases and peroxidases using a PCR and cloning approach with basidiomycete-specific primers. Between two and five laccase genes were detected in each strain. In contrast, only one gene encoding manganese peroxidase or versatile peroxidase was detected in each strain. The translated laccases and peroxidases amino acid sequences have not been described before. Extracellular crude enzymatic extracts produced by the UH strains, were able to degrade model chromophoric compounds such as anthraquinone and azo dyes. These findings hold promises for the development of a practical application for the treatment of textile industry wastewaters and also for bioremediation of polluted ecosystems by well-adapted native WRF strains.
白腐真菌(WRF)及其木质素分解酶(漆酶和过氧化物酶)被认为是从工业废水和受污染生态系统中去除与木质素相关的持久性有机污染物的有前途的生物技术工具。古巴已报道该属具有高度多样性;尽管如此,木质素分解酶及其基因的多样性仍未得到探索。在本研究中,从哈瓦那(古巴)城市生态系统中的腐朽木材中分离出13株本地WRF菌株。使用基于ITS序列的多重聚合酶链反应(PCR)方法将所有菌株鉴定为 种。所有 种菌株产生漆酶的水平高于非特异性过氧化物酶。细胞外酶提取物的天然聚丙烯酰胺凝胶电泳显示菌株之间漆酶同工酶模式具有高度多样性,表明这些 菌株产生的漆酶中存在不同的氨基酸序列。我们使用针对担子菌的特异性引物,通过PCR和克隆方法确定了编码漆酶和过氧化物酶的基因的多样性。每个菌株中检测到2至5个漆酶基因。相比之下,每个菌株中仅检测到一个编码锰过氧化物酶或多功能过氧化物酶的基因。之前尚未描述过翻译后的漆酶和过氧化物酶氨基酸序列。由 UH菌株产生的细胞外粗酶提取物能够降解模型发色化合物,如蒽醌和偶氮染料。这些发现为开发处理纺织工业废水的实际应用以及通过适应性良好的本地WRF菌株对污染生态系统进行生物修复带来了希望。