Mazzei Rosalinda, Bazzarelli Fabio, Terholsen Henrik, Nardi Monica, Piacentini Emma, Procopio Antonio, Bornscheuer Uwe T, Giorno Lidietta
Institute on Membrane Technology, National Research Council, CNR-ITM, University of Calabria, via P. Bucci, 17/C, 87036, Rende (Cosenza), Italy.
Department of Biotechnology and Enzyme Catalysis, Institute of Biochemistry, University of Greifswald, Felix-Hausdorff- Straße 4, 17487, Greifswald, Germany.
ChemSusChem. 2025 Mar 3;18(5):e202401707. doi: 10.1002/cssc.202401707. Epub 2024 Nov 13.
An integrated system of three membrane bioreactors (MBRs) has been developed that cascades three different enzymatic reactions. The integrated system was applied to produce hydroxytyrosol acetate from oleuropein extracted from olive leaves. Different reactor configurations for each reaction were tested and individually optimized to select the MBR to ensure high conversion and continuous production of oleuropein aglycone (OA), hydroxytyrosol (HY) and hydroxytyrosol acetate (HA). Based on this study, the most performing configuration of the integrated system was identified. In the first reaction, oleuropein was converted to OA using a biocatalytic membrane reactor (BMR) with immobilized β-glucosidase in polymeric membranes (conversion 95 %). The OA was then fed to another BMR, where it was converted to HY (conversion: 70 %) by an immobilized mutant of the promiscuous hydrolase/acyltransferase (PestE) (from the thermophilic archaeon Pyrobaculum calidifontis VA1). The HY produced was then acetylated using PestE immobilized on magnetic nanoparticles in a multiphase MBR (conversion: 98 %) and simultaneously extracted (extraction: 98 %) in ethyl acetate. The work demonstrates that continuous cascade enzymatic reactions can be engineered using artificial membranes to tailor enzyme compartmentalization, mass transport and phase contact according to reaction requirements. Besides, environmental factors proved the sustainability of the integrated membrane bioreactive system.
已开发出一种由三个膜生物反应器（MBR）组成的集成系统，该系统可实现三种不同的酶促反应串联。该集成系统用于从橄榄叶中提取的橄榄苦苷生产羟基酪醇乙酸酯。对每个反应的不同反应器配置进行了测试，并分别进行了优化，以选择MBR，以确保橄榄苦苷苷元（OA）、羟基酪醇（HY）和羟基酪醇乙酸酯（HA）的高转化率和连续生产。基于这项研究，确定了集成系统的最佳性能配置。在第一个反应中，使用在聚合物膜中固定化β-葡萄糖苷酶的生物催化膜反应器（BMR）将橄榄苦苷转化为OA（转化率95%）。然后将OA进料到另一个BMR中，在那里它被混杂水解酶/酰基转移酶（PestE）（来自嗜热古菌嗜热栖热袍菌VA1）的固定化突变体转化为HY（转化率：70%）。然后，使用固定在磁性纳米颗粒上的PestE在多相MBR中将产生的HY乙酰化（转化率：98%），并同时在乙酸乙酯中萃取（萃取率：98%）。这项工作表明，可以使用人工膜设计连续的级联酶促反应，以根据反应要求调整酶的分区、传质和相接触。此外，环境因素证明了集成膜生物反应系统的可持续性。
