Koplányi Gábor, Bell Evelin, Molnár Zsófia, Katona Gábor, Lajos Neumann Péter, Ender Ferenc, Balogh György T, Žnidaršič-Plazl Polona, Poppe László, Balogh-Weiser Diána
Department of Organic Chemistry and Technology, Budapest University of Technology and Economics, 1111, Műegyetem rkp. 3., Budapest, Hungary.
Institute of Enzymology, ELKH Research Center of Natural Sciences, 1117, Magyar tudosók krt. 2. Budapest, Hungary.
Chembiochem. 2023 Apr 3;24(7):e202200713. doi: 10.1002/cbic.202200713. Epub 2023 Mar 2.
The increasing application of recombinant enzymes demands not only effective and sustainable fermentation, but also highly efficient downstream processing and further stabilization of the enzymes by immobilization. In this study, a novel approach for the isolation and immobilization of His-tagged transaminase from Chromobacterium violaceum (CvTA) has been developed. A recombinant of CvTA was simultaneously isolated and immobilized by binding on silica nanoparticles (SNPs) with metal affinity linkers and additionally within poly(lactic acid) (PLA) nanofibers. The linker length and the nature of the metal ion significantly affected the enzyme binding efficiency and biocatalytic activity of CvTA-SNPs. The formation of PLA nanofibers by electrospinning enabled rapid embedding of CvTA-SNPs biocatalysts and ensured enhanced stability and activity. The developed advanced immobilization method reduces the time required for enzyme isolation, purification and immobilization by more than fourfold compared to a classical stepwise technique.
重组酶应用的不断增加不仅需要有效且可持续的发酵，还需要高效的下游加工以及通过固定化对酶进行进一步稳定化处理。在本研究中，已开发出一种从紫色色杆菌中分离和固定化His标签转氨酶（CvTA）的新方法。通过与金属亲和连接子结合在二氧化硅纳米颗粒（SNP）上，以及另外包埋在聚乳酸（PLA）纳米纤维内，同时实现了CvTA重组体的分离和固定化。连接子长度和金属离子性质显著影响CvTA-SNP的酶结合效率和生物催化活性。通过静电纺丝形成PLA纳米纤维能够快速包埋CvTA-SNP生物催化剂，并确保增强的稳定性和活性。与传统的逐步技术相比，所开发的先进固定化方法将酶分离、纯化和固定化所需的时间减少了四倍以上。
