Trametesversicolor是一种居住在木材中的琼脂菌,以其在硬木上引起强烈的白腐病腐烂的能力以及对酚类化合物的高耐受性而闻名。本工作的目的是深入了解这种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。对橡木上生长的杂色T.BRFM1218的分泌组的蛋白质组学分析揭示了一组200个分泌蛋白,其中包括染料脱色过氧化物酶TvDyP1和多功能过氧化物酶TvVP2。两种过氧化物酶都是在大肠杆菌中异源产生的,生化特征,并测试了氧化复杂底物的能力。发现两种过氧化物酶在酸性条件下对几种底物具有活性,TvDyP1在2.0至6.0的相对大的pH范围内非常稳定,而TvVP2仅在pH5.0至6.0下更稳定。还测试了两种酶的热稳定性,并且在全球范围内发现TvDyP1比TvVP2更稳定。在30至50°C的温度下孵育180分钟后,TvVP2的活性急剧下降,保留了10%至30%的初始活性。在同样的条件下,TvDyP1保留了其20-80%的酶活性。这两种蛋白质被催化表征,和TvVP2显示接受比TvDyP1更宽范围的还原底物。此外,两种酶都被发现对两种类黄酮有活性,槲皮素和儿茶素,在橡木中发现,其中TvVP2显示出两种化合物更快速的氧化。测试了它们对五种工业染料的脱色能力,与TvDyP1相比,TvVP2具有更大的氧化和脱色染料底物的能力。IMPORTANCETrametesversicolor是一种居住在木材中的琼脂菌,以其在硬木上引起强烈的白腐病腐烂的能力以及对酚类化合物的高耐受性而闻名。在白腐真菌中,杂色担子菌因其降解木材的能力而被广泛研究,特别是木质素,多亏了细胞外氧化酶系统.先前在经典木质素和锰过氧化物酶的几项工作中研究了相应的氧化系统,在这项研究中,杂色T.氧化系统的两个新成分,一种染料脱色过氧化物酶和一种多功能过氧化物酶,进行了深入的生化表征,并与其他真菌过氧化物酶进行了比较。
Trametesversicolor 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 T. versicolor BRFM 1218 grown on oak wood revealed a set of 200 secreted proteins, among which were the dye-decolorizing peroxidase TvDyP1 and the versatile peroxidase TvVP2. Both peroxidases were heterologously produced in Escherichia coli, 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 TvDyP1 was very stable over a relatively large pH range of 2.0 to 6.0, while TvVP2 was more stable at pH 5.0 to 6.0 only. The thermostability of both enzymes was also tested, and TvDyP1 was globally found to be more stable than TvVP2. After 180 min of incubation at temperatures ranging from 30 to 50°C, the activity of TvVP2 drastically decreased, with 10 to 30% of the initial activity retained. Under the same conditions, TvDyP1 retained 20 to 80% of its enzyme activity. The two proteins were catalytically characterized, and TvVP2 was shown to accept a wider range of reducing substrates than TvDyP1. Furthermore, both enzymes were found to be active against two flavonoids, quercetin and catechin, found in oak wood, with TvVP2 displaying more rapid oxidation of the two compounds. They were tested for the ability to decolorize five industrial dyes, and TvVP2 presented a greater ability to oxidize and decolorize the dye substrates than TvDyP1.IMPORTANCETrametesversicolor 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 T. versicolor 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 T. versicolor, one dye-decolorizing peroxidase and one versatile peroxidase, were biochemically characterized in depth and compared to other fungal peroxidases.