We herein describe a bioinspired solid-phase assembly of a multienzyme system scaffolded on an artificial cellulosome. An alcohol dehydrogenase and an ω-transaminase were fused to cohesin and dockerin domains to drive their sequential and ordered coimmobilization on agarose porous microbeads. The resulting immobilized scaffolded enzymatic cellulosome was characterized through quartz crystal microbalance with dissipation and confocal laser scanning microscopy to demonstrate that both enzymes interact with each other and physically colocalize within the microbeads. Finally, the assembled multifunctional heterogeneous biocatalyst was tested for the one-pot conversion of alcohols into amines. By using the physically colocalized enzymatic system confined into porous microbeads, the yield of the corresponding amine was 1.3 and 10 times higher than the spatially segregated immobilized system and the free enzymes, respectively. This work establishes the basis of a new concept to organize multienzyme systems at the nanoscale within solid and porous immobilization carriers.

中文翻译：

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多酶系统固相组装成人工纤维素体

我们在此描述了在人工纤维素体上支架的多酶系统的仿生固相组装。醇脱氢酶和 ω-转氨酶与 cohesin 和 dockerin 结构域融合，以驱动它们在琼脂糖多孔微珠上的顺序和有序共固定。由此产生的固定支架酶纤维素小体通过石英晶体微天平与耗散和共聚焦激光扫描显微镜进行表征，以证明两种酶相互作用并在微珠内物理共定位。最后，测试组装的多功能多相生物催化剂将醇一锅法转化为胺。通过使用限制在多孔微珠中的物理共定位酶系统，相应胺的产率为 1。分别比空间隔离固定系统和游离酶高 3 倍和 10 倍。这项工作为在固体和多孔固定载体内在纳米尺度上组织多酶系统的新概念奠定了基础。