Search for natural laccase mediator and its application for green chemistry

漆酶-介体体系是木质素降解能力最强的体系之一。然而，从木质素降解真菌培养中提取的天然漆酶介体尚未见报道。为了解决这个问题，我们考虑了用于搜索介体的检测系统。首先，我们建立了一套用漆酶-介体体系处理的未漂硫酸盐浆的破损能力(白度和卡伯值)的测定系统。然而，由于操作复杂，我们拒绝了这一系统。其次，我们设计了一套基于蒽氧化的检测系统。漆酶只有在介体1-羟基苯并三氮唑(HBT)存在的情况下才能氧化菲。以杂色曲霉(Trametes versicolor漆酶，Fr.)为介体，制备了超滤渗透液。B)和其真菌水提物(Fr.f)。它们都能作为漆酶的中介体(降解率，F：67.3%，B：75.5%)。然后，这些提取物被分成正己烷可溶(Hex)和乙酸乙酯可溶(EtOAc)。其中3个组分具有介体活性(降解率，F-Hex：70.0%，B-Hex：70.0%，B-EtOAc：75.6%)。在这些抽象体中，调解人的存在被认为是很有可能的。

Laccase-mediator system is one of the most powerful lignin degrading systems. However, the natural laccase mediator from the culture of lignin-degrading fungi has been not reported yet. To address this question, we considered the assay systems for searching the mediator. First, we developed an assay system using the breaching abilities (brightness and Kappa number) of unbleached kraft pulp treated with laccase-mediator system. However, we rejected this system for the complicated operation. Second, we planed an assay system by using of anthracene oxidation. Laccase could only oxidized anthracene in the presence of mediator, for example 1-hydroxybenzotriazole (HBT). Consequently, this system was simple and quick assay system by using HPLC.For the mediator candidates, we prepared the ultrafiltration permeate of Trametes versicolor laccase (Fr. B) and water extracts of its fungus (Fr. F). They could act as the laccase mediator for anthracene oxidation (Degradation rate, F : 67.3%, B : 75.5 %). Then, these extracts were fractionated to hexane-soluble (Hex) and ethyl acetate-soluble (EtOAc). Three fractions of them have mediator activities (Degradation rate, F-Hex : 70.0%, B-Hex : 70.0 %, B-EtOAc : 75.6 %). The existence of mediator in these extractives is seen as a strong possibility.