College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211816, China.
School of Food Science and Pharmaceutical Engineering, Nanjing Normal University, Nanjing 210023, China.
ACS Appl Mater Interfaces. 2023 Apr 26;15(16):19807-19816. doi: 10.1021/acsami.2c22094. Epub 2023 Mar 17.
The multienzyme cascade has received growing attention to obtain structurally defined glycans . However, due to poor enzyme stability and low compatibility between glycoenzymes, artificially designed multienzyme pathways to access glycans are often inefficient. Herein, based on the strategy "Modular-Enzymes Assembly by Spatial Segregation" (MASS), we developed a universal immobilization platform to assemble multiple glycoenzymes in compartmentalized MOF particles, inside and outside, significantly reducing the undesired interference and cross-inhibitions. By changing the enzyme modules, a series of glycosyl donor, disaccharides, oligosaccharides, and polysaccharides bearing cofactor regeneration were efficiently prepared. This bioreactor was further successfully applied to the reaction system with high substrate concentration to demonstrate its industrial potential. This robust multienzyme immobilization platform should serve to promote the enzymatic synthesis of more complex glycans.
多酶级联反应受到越来越多的关注，以获得结构定义明确的聚糖。然而，由于酶稳定性差和糖基酶之间的兼容性低，人为设计的获得聚糖的多酶途径往往效率低下。在此，基于“空间隔离的模块化酶组装”（MASS）策略，我们开发了一种通用的固定化平台，将多个糖基酶组装在分隔的 MOF 颗粒内、内外部分，显著减少了不必要的干扰和交叉抑制。通过改变酶模块，高效制备了一系列带有辅因子再生的糖基供体、二糖、寡糖和多糖。该生物反应器进一步成功应用于高底物浓度的反应体系，展示了其工业潜力。这种稳健的多酶固定化平台有望促进更复杂聚糖的酶促合成。
