Coşkun Banuay, Say Rıdvan, Ersöz Arzu
Eskişehir Technical University, Faculty of Science, Department of Chemistry, Eskişehir, Türkiye.
Bionkit Co. Ltd. Anadolu University Teknopark, Eskişehir, Türkiye.
Int J Biol Macromol. 2025 Aug 29;327(Pt 1):147249. doi: 10.1016/j.ijbiomac.2025.147249.
This study introduces a reusable cryogenic column system immobilized with laccase enzyme (laccase-M) for the sustainable synthesis of a novel tyrosine-based crosslinker, dityrosine-dimethacrylate (Dityr-diMA). By leveraging methacrylated tyrosine (Tyr-MA) as a biocatalytic platform, the laccase-M column enables efficient polymerization via oxidative coupling, eliminating the need for toxic photoinitiators or metal catalysts. Compared to conventional HEMA-based columns, the laccase-M system achieved a 19.5% higher water retention capacity (819.83% → 980.16% swelling ratio) and maintained 74.02% relative enzyme activity (172.32 → 127.55μMmin) over 50 reuse cycles, with a remarkably low decay rate of 0.52 % per cycle, demonstrating unprecedented operational stability for cryo-immobilized laccase systems. The enzymatic method also reduced organic solvent consumption by 60 %. Structural analyses (SEM, BET) revealed that the crosslinked Dityr-diMA network exhibits a homogeneous pore architecture (BET surface area: 6.219mg) and enhanced material stability. Kinetic studies confirmed the system's superior substrate affinity, with a 10-fold decrease in Km values (0.0192 → 0.0021 mM), highlighting its industrial scalability. Compared to UV crosslinking methods (Eosin Y, Irgacure 2959, LAP) and (metal complexes ([Ru(II)(bpy)])), the enzymatic approach provided a controlled polymerization mechanism with low cytotoxicity. This study demonstrates with experimental data that enzymatic crosslinking can be used as a non-toxic, scalable, and efficient strategy in the design of high-performance biomaterials; It offers an environmentally friendly alternative to traditional methods, paving the way for safer and more sustainable approaches in biomedical and tissue engineering applications.
本研究介绍了一种固定有漆酶（漆酶-M）的可重复使用的低温柱系统，用于可持续合成一种新型的基于酪氨酸的交联剂二酪氨酸二甲基丙烯酸酯（Dityr-diMA）。通过利用甲基丙烯酸化酪氨酸（Tyr-MA）作为生物催化平台，漆酶-M柱能够通过氧化偶联实现高效聚合，无需使用有毒的光引发剂或金属催化剂。与传统的基于甲基丙烯酸羟乙酯（HEMA）的柱相比，漆酶-M系统的保水能力提高了19.5%（溶胀率从819.83%提高到980.16%），并且在50次重复使用循环中保持了74.02%的相对酶活性（从172.32降至127.55μMmin），每个循环的衰减率极低，仅为0.52%，这表明低温固定化漆酶系统具有前所未有的操作稳定性。酶法还使有机溶剂消耗量减少了60%。结构分析（扫描电子显微镜、比表面积测定）表明，交联的Dityr-diMA网络具有均匀的孔结构（比表面积：6.219mg），且材料稳定性增强。动力学研究证实了该系统具有优异的底物亲和力，米氏常数（Km）值降低了10倍（从0.0192降至0.0021mM），突出了其工业可扩展性。与紫外线交联方法（曙红Y、光引发剂Irgacure 2959、光引发剂LAP）以及（金属配合物（[Ru(II)(bpy)]））相比，酶法提供了一种具有低细胞毒性的可控聚合机制。本研究通过实验数据表明，酶交联可作为一种无毒、可扩展且高效的策略用于高性能生物材料的设计；它为传统方法提供了一种环境友好的替代方案，为生物医学和组织工程应用中更安全、更可持续的方法铺平了道路。
