Spano Michael B, Tran Brandan H, Majumdar Sudipta, Weiss Gregory A
Department of Chemistry, University of California, Irvine, Irvine, California 92697-2025, United States.
Department of Molecular Biology and Biochemistry, University of California, Irvine, Irvine, California 92697-3900, United States.
J Org Chem. 2020 Jul 2;85(13):8480-8488. doi: 10.1021/acs.joc.0c00789. Epub 2020 Jun 17.
In continuous flow biocatalysis, chemical transformations can occur under milder, greener, more scalable, and safer conditions than conventional organic synthesis. However, the method typically involves extensive screening to optimize each enzyme's immobilization on its solid support material. The task of weighing solids for large numbers of experiments poses a bottleneck for screening enzyme immobilization conditions. For example, screening conditions often require multiple replicates exploring different support chemistries, buffer compositions, and temperatures. Thus, we report 3D-printed labware designed to measure and handle solids in multichannel format and expedite screening of enzyme immobilization conditions. To demonstrate the generality of these advances, alkaline phosphatase, glucose dehydrogenase, and laccase were screened for immobilization efficiency on seven resins. The results illustrate the requirements for optimization of each enzyme's loading and resin choice for optimal catalytic performance. Here, 3D-printed labware can decrease the requirements for an experimentalist's time by >95%. The approach to rapid optimization of enzyme immobilization is applicable to any enzyme and many solid support resins. Furthermore, the reported devices deliver precise and accurate aliquots of essentially any granular solid material.
在连续流动生物催化中,与传统有机合成相比,化学反应可以在更温和、更环保、更具可扩展性和更安全的条件下进行。然而,该方法通常需要进行大量筛选,以优化每种酶在其固体载体材料上的固定化。在大量实验中称量固体的任务成为筛选酶固定化条件的瓶颈。例如,筛选条件通常需要多次重复实验,探索不同的载体化学性质、缓冲液组成和温度。因此,我们报告了一种3D打印实验室器具,其设计用于以多通道形式测量和处理固体,并加快酶固定化条件的筛选。为了证明这些进展的通用性,我们对碱性磷酸酶、葡萄糖脱氢酶和漆酶在七种树脂上的固定化效率进行了筛选。结果说明了优化每种酶的负载量和树脂选择以实现最佳催化性能的要求。在此,3D打印实验室器具可以将实验人员的时间需求减少95%以上。这种快速优化酶固定化的方法适用于任何酶和许多固体载体树脂。此外,所报道的装置能够精确且准确地分配基本上任何颗粒状固体材料。