INRA, Aix-Marseille Université, UMR1163, Biodiversité et Biotechnologie Fongiques, Marseille, France
INRA, UMR 1136, Interactions Arbres/Micro-Organismes, Champenoux, France.
Appl Environ Microbiol. 2018 Apr 2;84(8). doi: 10.1128/AEM.02826-17. Print 2018 Apr 15.
is a wood-inhabiting agaricomycete known for its ability to cause strong white-rot decay on hardwood and for its high tolerance of phenolic compounds. The goal of the present work was to gain insights into the molecular biology and biochemistry of the heme-including class II and dye-decolorizing peroxidases secreted by this fungus. Proteomic analysis of the secretome of BRFM 1218 grown on oak wood revealed a set of 200 secreted proteins, among which were the dye-decolorizing peroxidase DyP1 and the versatile peroxidase VP2. Both peroxidases were heterologously produced in , biochemically characterized, and tested for the ability to oxidize complex substrates. Both peroxidases were found to be active against several substrates under acidic conditions, and DyP1 was very stable over a relatively large pH range of 2.0 to 6.0, while VP2 was more stable at pH 5.0 to 6.0 only. The thermostability of both enzymes was also tested, and DyP1 was globally found to be more stable than VP2. After 180 min of incubation at temperatures ranging from 30 to 50°C, the activity of VP2 drastically decreased, with 10 to 30% of the initial activity retained. Under the same conditions, DyP1 retained 20 to 80% of its enzyme activity. The two proteins were catalytically characterized, and VP2 was shown to accept a wider range of reducing substrates than DyP1. Furthermore, both enzymes were found to be active against two flavonoids, quercetin and catechin, found in oak wood, with VP2 displaying more rapid oxidation of the two compounds. They were tested for the ability to decolorize five industrial dyes, and VP2 presented a greater ability to oxidize and decolorize the dye substrates than DyP1. is a wood-inhabiting agaricomycete known for its ability to cause strong white-rot decay on hardwood and for its high tolerance of phenolic compounds. Among white-rot fungi, the basidiomycete has been extensively studied for its ability to degrade wood, specifically lignin, thanks to an extracellular oxidative enzymatic system. The corresponding oxidative system was previously studied in several works for classical lignin and manganese peroxidases, and in this study, two new components of the oxidative system of , one dye-decolorizing peroxidase and one versatile peroxidase, were biochemically characterized in depth and compared to other fungal peroxidases.
是一种木质腐生伞菌，以其对硬木的强烈白腐能力和对酚类化合物的高耐受性而闻名。本研究的目的是深入了解该真菌分泌的血红素包括 II 类和染料脱色过氧化物酶的分子生物学和生物化学。BRFM 1218 在橡木上生长时的分泌组的蛋白质组学分析揭示了一组 200 种分泌蛋白，其中包括染料脱色过氧化物酶 DyP1 和多功能过氧化物酶 VP2。这两种过氧化物酶均在 中异源表达，进行了生化表征，并测试了其氧化复杂底物的能力。在酸性条件下，发现这两种过氧化物酶都能有效地氧化几种底物，DyP1 在相对较大的 pH 2.0 至 6.0 范围内非常稳定，而 VP2 仅在 pH 5.0 至 6.0 时更稳定。还测试了这两种酶的热稳定性，发现 DyP1 总体上比 VP2 更稳定。在 30 至 50°C 的温度范围内孵育 180 分钟后，VP2 的活性急剧下降，保留了初始活性的 10 至 30%。在相同条件下，DyP1 保留了 20 至 80%的酶活性。对这两种蛋白质进行了催化特性分析，结果表明 VP2 能接受比 DyP1 更广泛的还原底物。此外，发现这两种酶都能有效地氧化两种在橡木中发现的类黄酮，槲皮素和儿茶素，VP2 对这两种化合物的氧化速度更快。还测试了它们对五种工业染料的脱色能力，VP2 比 DyP1 具有更强的氧化和脱色染料底物的能力。是一种木质腐生伞菌，以其对硬木的强烈白腐能力和对酚类化合物的高耐受性而闻名。在白腐真菌中，担子菌由于其具有木质素，特别是木质素的体外氧化酶系统，已被广泛研究。在之前的几项研究中，对该氧化系统的相应氧化系统进行了研究，包括经典木质素和锰过氧化物酶，在本研究中，深入研究了木质素的两种新的氧化系统成分，一种是染料脱色过氧化物酶，另一种是多功能过氧化物酶，并与其他真菌过氧化物酶进行了比较。
