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In this study, a new pathway with four enzymes including formate dehydrogenase (PaFDH), formaldehyde dehydrogenase (BmFADH), glycolaldehyde synthase (PpGALS), and alcohol dehydrogenase (GoADH) is developed for directly converting CO<jats:sub>2<\/jats:sub> gas molecules to ethylene glycol (EG) in vitro. A rhodium\u2010based NADH regeneration electrode is constructed to continuously provide the proton and electron of this multi\u2010enzymatic cascade reaction. The prepared electrode can reach the Faradaic Efficiency (FE) of 82.9% at \u22120.6\u00a0V (vs. Ag\/AgCl) and the NADH productivity of 0.737\u00a0mM h<jats:sup>\u22121<\/jats:sup>. Shortening the reaction path is crucial for multi\u2010enzymatic cascade reactions. Here, a hydrogen\u2010bonded organic framework (HOF) nano\u2010reactor is successfully developed to immobilize four enzymes in one pot with a striking enzyme loading capacity (990\u00a0mg enzyme\u00a0g<jats:sup>\u22121<\/jats:sup> material). Through integrating and optimization of NADH electro\u2010regeneration and enzymatic catalysis in one pot, 0.15\u00a0mM EG is achieved with an average conversion rate of 7.15 \u00d7 10<jats:sup>\u22127<\/jats:sup>\u00a0mmol CO<jats:sub>2<\/jats:sub> min<jats:sup>\u22121<\/jats:sup> mg<jats:sup>\u22121<\/jats:sup> enzymes in 6\u00a0h. These results shed light on electro\u2010driven multi\u2010enzymatic cascade conversion of C\u2500C coupling from CO<jats:sub>2<\/jats:sub> in the nano\u2010reactor.<\/jats:p>","DOI":"10.1002\/advs.202407204","type":"journal-article","created":{"date-parts":[[2024,9,4]],"date-time":"2024-09-04T15:29:39Z","timestamp":1725463779000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":26,"title":["Electro\u2010Driven Multi\u2010Enzymatic Cascade Conversion of CO<sub>2<\/sub> to Ethylene Glycol in Nano\u2010Reactor"],"prefix":"10.1002","volume":"11","author":[{"given":"Likun","family":"Luan","sequence":"first","affiliation":[{"name":"Beijing Key Laboratory of Ionic Liquids Clean Process CAS Key Laboratory of Green Process and Engineering State Key Laboratory of Mesoscience and Engineering Institute of Process Engineering Chinese Academy of Sciences  Beijing 100190 P. R. 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