Lignin is a major byproduct of pulp and paper industries, which is resistant to depolymerization due to its heterogeneous structure. The enzymes peroxidases can be utilized as potent bio-catalysts to degrade lignin. In the current study, an Efeb gene of 1251bp encoding DyP-type peroxidase from Bacillus sp. strain BL5 (DyPBL5) was amplified, cloned into a pET-28a (+) vector and expressed in Escherichia coli BL21 (DE3) cells. A 46 kDa protein of DyPBL5 was purified through ion-exchange chromatography. Purified DyPBL5 was active at wide temperature (25-50 °C) and pH (3.0-8.0) range with optimum activity at 35 °C and pH 5.0. Effects of different chemicals on DyPBL5 were determined. The enzyme activity was strongly inhibited by SDS, DDT and β-mercaptoethanol, whereas stimulated in the presence of organic solvents such as methanol and ethanol. The kinetic parameters were determined and K_m, V_max and K_cat values were 1.06 mM, 519.751/4mol/min/mg and 395 S̶ ¹, respectively. Docking of DyPBL5 with ABTS revealed that, Asn 244, Arg 339, Asp 383 and Thr 389 are putative amino acids, taking part in the oxidation of ABTS. The recombinant DyPBL5 resulted in the reduction of lignin contents up to 26.04%. The SEM and FT-IR analysis of test samples gave some indications about degradation of lignin by DyPBL5. Various low molecular weight lignin degradation products were detected by analyzing the samples through gas chromatography mass spectrometry. High catalytic efficiency and lignin degradation rate make DyPBL5 an ideal bio-catalyst for remediation of lignin-contaminated sites.

木质素是纸浆和造纸工业的主要副产品，由于其异质结构而难以解聚。过氧化物酶可用作降解木质素的有效生物催化剂。在目前的研究中，1251bp 的 Efeb 基因编码来自芽孢杆菌的DyP 型过氧化物酶。菌株 BL5 (DyPBL5) 被扩增，克隆到 pET-28a (+) 载体并在大肠杆菌中表达BL21 (DE3) 细胞。DyPBL5 的 46 kDa 蛋白质通过离子交换色谱法纯化。纯化的 DyPBL5 在较宽的温度 (25-50 °C) 和 pH (3.0-8.0) 范围内具有活性，在 35 °C 和 pH 5.0 下具有最佳活性。确定了不同化学品对 DyPBL5 的影响。SDS、DDT 和β-巯基乙醇对酶活性有强烈抑制作用，而在有机溶剂如甲醇和乙醇存在下则受到刺激。动力学参数进行测定和ķ_米，V_最大和ķ _{CA Ť}值分别为1.06毫米，519.751 / 4摩尔/分钟/毫克和395š ¹， 分别。DyPBL5 与 ABTS 的对接表明，Asn 244、Arg 339、Asp 383 和 Thr 389 是推定的氨基酸，参与了 ABTS 的氧化。重组 DyPBL5 导致木质素含量减少高达 26.04%。测试样品的 SEM 和 FT-IR 分析给出了一些关于 DyPBL5 降解木质素的迹象。通过气相色谱质谱法分析样品，检测到各种低分子量木质素降解产物。高催化效率和木质素降解率使 DyPBL5 成为修复木质素污染场地的理想生物催化剂。