Green\nand Fast Synthesis of NiCo-MOF for Simultaneous\nPurification–Immobilization of Bienzyme to Catalyze the Synthesis\nof Ginsenoside Rh2

Traditional\nmetal–organic frameworks (MOFs) preparation\nis generally time-consuming, polluting, and lacking specificity for\nenzyme immobilization. This paper introduced a facile, rapid, and\ngreen method to produce three MOFs subsequently employed to purify\nand coimmobilize recombinant glycosyltransferase (UGT) and recombinant\nsucrose synthetase (SUSy) using histidine tag (His-tag) for the specific\nadsorption of Ni<sup>2+</sup> and Co<sup>2+</sup> from MOFs. This\nmethod simplified enzyme purification from crude extracts and enabled\nenzymes to be reused. The results demonstrated that NiCo-MOF exhibited\na higher enzyme load (115.9 mg/g) than monometallic MOFs. Additionally,\nthe NiCo-MOF@UGT&SUSy demonstrated excellent stability and efficiently\nproduced the rare ginsenoside Rh2 by catalyzing a coupling reaction\n(95.6 μg/mL), solving the problem of the substrate cost of uridine\ndiphosphate glucose (UDPG). The NiCo-MOF@UGT&SUSy retained 68.97%\nof the initial activity after 10 cycles. Finally, molecular docking\nstudies elucidated the conversion mechanism of the target product\nRh2. This technique is important in the industrialization of ginsenoside\nproduction and enzyme purification.