Trubitsina Liubov, Egorov Konstantin, Abdullatypov Azat, Petrakova Marina, Trubitsin Ivan, Alferov Sergey, Leontievsky Alexey, Ponamoreva Olga
G.K. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Federal Research Center "Pushchino Scientific Center for Biological Research of the Russian Academy of Sciences", 142290 Pushchino, Russia.
Department of Biotechnology, Tula State University, 300012 Tula, Russia.
Int J Mol Sci. 2025 Sep 19;26(18):9143. doi: 10.3390/ijms26189143.
A novel two-domain small laccasefrom (SoSL) was produced through recombination in and purified by affinity chromatography. The properties (thermal optimum and thermostability, pH optima and pH-stability), kinetic characteristics, substrate specificity and dye decolorization ability were estimated. Laccase SoSL was able to oxidize 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS)and 2,6-dimethoxyphenol (2,6-DMP) with at a maximal rate at pH 3.5 and 9.0, respectively, and was stable at pH 9.0 (retained 75% activity after incubation at room temperature for 120 h). High enzyme affinity to ABTS is caused by an expanded area occupied by aromatic amino acid residues on its surface. Substrate-directed immobilization of the enzyme was performed using naphthylated multiwalled carbon nanotubes (MWCNTs), and a high oxygen reduction reaction potential (+0.62 V vs. normal hydrogen electrode (NHE)) was observed. The above-mentioned features make this enzyme a promising one for further studies in bioremediation and biological fuel cell technologies.
通过在大肠杆菌中重组表达并经亲和层析纯化，获得了一种新型的双结构域小漆酶（SoSL）。对其性质（最适温度和热稳定性、最适pH和pH稳定性）、动力学特征、底物特异性和染料脱色能力进行了评估。漆酶SoSL能够分别在pH 3.5和9.0时以最大速率氧化2,2'-联氮-双（3-乙基苯并噻唑啉-6-磺酸）（ABTS）和2,6-二甲氧基苯酚（2,6-DMP），并且在pH 9.0时稳定（室温孵育120小时后保留75%的活性）。其对ABTS的高酶亲和力是由其表面芳香族氨基酸残基占据的扩展区域引起的。使用萘基化多壁碳纳米管（MWCNT）对该酶进行了底物导向固定化，并观察到高氧还原反应电位（相对于标准氢电极（NHE）为+0.62 V）。上述特性使这种酶在生物修复和生物燃料电池技术的进一步研究中具有广阔前景。
