Laccases are a class of multi-copper oxidases with important industrial values. A thermotolerant laccase produced by a basidiomycete fungal strain Cerrena unicolor CGMCC 5.1011 was studied. With glycerin and peptone as the carbon and nitrogen sources, respectively, a maximal laccase activity of 121.7 U/mL was attained after cultivation in the shaking flask for 15 days. Transcriptomics analysis revealed an expressed laccase gene family of 12 members in C. unicolor strain CGMCC 5.1011, and the gene and cDNA sequences were cloned. A glycosylated laccase was purified from the fermentation broth of Cerrena unicolor CGMCC 5.1011 and corresponded to Lac2 based on MALDI-TOF MS/MS identification. Lac2 was stable at pH 5.0 and above, and was resistant to organic solvents. Lac2 displayed remarkable thermostability, with half-life time of 1.67 h at 70 ºC. Consistently, Lac2 was able to completely decolorize malachite green (MG) at high temperatures, whereas Lac7 from Cerrena sp. HYB07 resulted in accumulation of colored MG transformation intermediates. Molecular dynamics simulation of Lac2 was conducted, and possible mechanisms underlying Lac2 thermostability were discussed. The robustness of C. unicolor CGMCC 5.1011 laccase would not only be useful for industrial applications, but also provide a template for future work to develop thermostable laccases.

漆酶是一类具有重要工业价值的多铜氧化酶。研究了由担子菌真菌Cerrena unicolor CGMCC 5.1011产生的耐热漆酶。用甘油和蛋白p分别作为碳和氮源，在摇瓶中培养15天后，最大漆酶活性达到121.7 U / mL。转录组学分析揭示了表达的漆酶基因家族，其由C. unicolor菌株CGMCC 5.1011中的12个成员组成，并克隆了基因和cDNA序列。从Cerrena unicolor的发酵液中纯化糖基化的漆酶CGMCC 5.1011，对应于基于MALDI-TOF MS / MS识别的Lac2。Lac2在pH 5.0和更高的条件下稳定，并且对有机溶剂具有抵抗力。Lac2具有出色的热稳定性，在70ºC下的半衰期为1.67 h。一致地，Lac2能够在高温下完全使孔雀石绿（MG）脱色，而来自Cerrena sp。的Lac7。HYB07导致有色MG转化中间体积累。进行了Lac2的分子动力学模拟，并讨论了Lac2热稳定性的潜在机理。C. unicolor CGMCC 5.1011漆酶的坚固性不仅可用于工业应用，而且还为开发耐高温漆酶的未来工作提供了模板。