University of Belgrade, Faculty of Chemistry, Studentski trg 16, P.O. Box 51, Belgrade, 11158, Serbia.
Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Vojvode Stepe 444a, Belgrade, 11000, Serbia.
Enzyme Microb Technol. 2020 Jan;132:109411. doi: 10.1016/j.enzmictec.2019.109411. Epub 2019 Aug 20.
Biocatalytic oxidations mediated by laccases are gaining importance due to their versatility and beneficial environmental effects. In this study, the oxidation of 1,4-dihydropyridines has been performed using three different types of bacterial laccase-based catalysts: purified laccase from Bacillus licheniformis ATCC 9945a (BliLacc), Escherichia coli whole cells expressing this laccase, and bacterial nanocellulose (BNC) supported BliLacc catalysts. The catalysts based on bacterial laccase were compared to the commercially available Trametes versicolor laccase (TvLacc). The oxidation product of 2,6-dimethyl-1,4-dihydropyridine-3,5-dicarboxylate was obtained within 7-24 h with good yields (70-99%) with all three biocatalysts. The substrate scope was examined with five additional 1,4-dihydropyridines, one of which was oxidized in high yield. Whole-cell biocatalyst was stable when stored for up to 1-month at 4 °C. In addition, evidence has been provided that multicopper oxidase CueO from the E. coli expression host contributed to the oxidation efficiency of the whole-cell biocatalyst. The immobilized whole-cell biocatalyst showed satisfactory activity and retained 37% of its original activity after three biotransformation cycles.
由于其多功能性和有益的环境效应，漆酶介导的生物催化氧化受到越来越多的关注。在这项研究中，使用三种不同类型的细菌漆酶基催化剂进行了 1,4-二氢吡啶的氧化：来自地衣芽孢杆菌 ATCC 9945a 的纯化漆酶（BliLacc）、表达这种漆酶的大肠杆菌全细胞和细菌纳米纤维素（BNC）支持的 BliLacc 催化剂。将细菌漆酶基催化剂与市售的云芝漆酶（TvLacc）进行了比较。用所有三种生物催化剂在 7-24 小时内获得了 2,6-二甲基-1,4-二氢吡啶-3,5-二羧酸酯的氧化产物，产率良好（70-99%）。用另外五种 1,4-二氢吡啶检查了底物范围，其中一种以高产率被氧化。当在 4°C 下储存长达 1 个月时，全细胞生物催化剂是稳定的。此外，有证据表明，来自大肠杆菌表达宿主的多铜氧化酶 CueO 有助于全细胞生物催化剂的氧化效率。固定化全细胞生物催化剂表现出令人满意的活性，在三个生物转化循环后保留了其原始活性的 37%。
