Institute of Water Research, University of Granada, Ramón y Cajal, Fray Luís 4, Granada 18071, Spain.
Institute of Water Research, University of Granada, Ramón y Cajal, Fray Luís 4, Granada 18071, Spain; Department of Microbiology, Pharmacy Faculty, University of Granada, Spain.
Ecotoxicol Environ Saf. 2018 Sep 15;159:127-135. doi: 10.1016/j.ecoenv.2018.04.051. Epub 2018 May 4.
Polycyclic aromatic hydrocarbons (PAHs) are widely distributed in polluted environments and are included in the priority list of toxic compounds. Previous studies have shown that the fungus Penicillium oxalicum, isolated from a hydrocarbon-polluted pond, has a great capability to transform different PAHs in short periods under submerged fermentation conditions. Although cytochrome p450s (CYPs) seems to be the main responsible enzyme in this process, changes in proteome profile remains poorly understood. The aim of this work was to characterise molecular disturbances in the cytosolic and microsomal sub-proteomes of P. oxalicum by applying two-dimensional (2D) gel electrophoresis and label-free quantitative proteomics during anthracene biodegradation. Our results showed that by using 2D-gels, 10 and 8 differential proteins were over-expressed in the cytosolic and microsomal fractions, respectively. Most of them were related to stress response. Shotgun proteomics allowed the identification of 158 and 174 unique protein species that differentially accumulated during anthracene biotransformation, such as CYPs, epoxide hydrolases and transferases enzymes, belonging to Phase I and Phase II of the metabolism of xenobiotics, contributing to the anthracene biodegradation pathway. These results confirm the biological significance of ascomycetes fungi the rol of CYPs on biodegradation and the need of a deeper knowledge on fungal proteomics for the application of the appropriate microorganisms in biodegradation processes.
多环芳烃（PAHs）广泛分布于污染环境中，被列入有毒化合物的优先清单。先前的研究表明，从烃污染池塘中分离出的青霉（Penicillium oxalicum）真菌在液体深层发酵条件下具有在短时间内转化不同 PAHs 的巨大能力。虽然细胞色素 P450s（CYPs）似乎是这个过程中的主要负责酶，但对蛋白质组谱的变化仍知之甚少。本研究的目的是通过应用二维（2D）凝胶电泳和无标记定量蛋白质组学在蒽生物降解过程中，研究草酸青霉的胞质和微粒体亚蛋白质组中的分子干扰。我们的结果表明，通过使用 2D 凝胶，在胞质和微粒体部分分别过表达了 10 种和 8 种差异蛋白。它们大多数与应激反应有关。鸟枪法蛋白质组学鉴定了 158 种和 174 种在蒽生物转化过程中差异积累的独特蛋白质，如细胞色素 P450s、环氧化物水解酶和转移酶酶，属于外源物代谢的 I 相和 II 相，有助于蒽的生物降解途径。这些结果证实了曲霉菌在生物降解中的生物学意义，CYPs 对生物降解的作用，以及为了在生物降解过程中应用适当的微生物，需要更深入地了解真菌蛋白质组学。
