Exploration of feruloyl esterase (FAE) with the resistance to heat and alkali conditions in biobleaching process to improve the separation efficiency of lignocellulose is the key to achieving green papermaking. Herein, we expressed FAEB of C. thermophilum and obtained a thermostable alkaline FAE that can effectively promote the removal of lignin from pulp. The faeB gene was successfully obtained through genomic Blast strategy and high-efficiency expressed under the control of strong alcohol oxidase promoter in Pichia pastoris. The recombinant CtFAEB has an optimal temperature of 65 °C and pH of 7.0. After treated at 65 °C for 1 h, CtFAEB can still retain 63.21 % of its maximum activity, showing a good thermal stability. In addition, the recombinant CtFAEB has broad pH stability and can retain about 56 % of the maximum activity even at pH 11.0. Compared with the effect of mesophilic FAE, pretreatment with thermostable CtFAEB can promote the delignification by laccase and alkaline hydrogen peroxide from the pulp at 70 °C and pH 9.0. Alignment of the protein sequences of CtFAEB and mesophilic FAE suggested that the percentage of amino acids that easily form alpha helix in CtFAEB increases, which enhances its structural rigidity and thereby improves its thermal stability and alkali tolerance. Our study provides an effective method to obtain thermostable and alkaline FAEs, which will promote its application in biobleaching and other biorefining industries.

探索在生物漂白过程中具有耐热和耐碱性条件的阿魏酸酯酶（FAE）以提高木质纤维素的分离效率是实现绿色造纸的关键。在此，我们表达了嗜热梭菌的 FAEB，获得了一种耐热的碱性 FAE，可以有效促进纸浆中木质素的去除。的FAE通过基因组高炉策略和高效率的强醇氧化酶启动子中的控制下表达被成功地获得的B基因毕赤酵母. 重组 CtFAEB 的最适温度为 65 °C，pH 值为 7.0。在 65°C 处理 1 h 后，CtFAEB 仍可保留其最大活性的 63.21%，显示出良好的热稳定性。此外，重组 CtFAEB 具有广泛的 pH 稳定性，即使在 pH 11.0 时也能保留约 56% 的最大活性。与嗜温 FAE 的效果相比，用耐热 CtFAEB 预处理可以促进漆酶和碱性过氧化氢在 70°C 和 pH 9.0 下从纸浆中脱木质素。CtFAEB和嗜温FAE的蛋白质序列比对表明，CtFAEB中容易形成α螺旋的氨基酸百分比增加，从而增强了其结构刚性，从而提高了其热稳定性和碱耐受性。我们的研究提供了一种获得耐热和碱性 FAE 的有效方法，