Carbon dioxide biomedication has attracted more research recently as it is a sustainable and environmentally friendly method of converting carbon into a variety of indispensable chemicals. However, studies on the development of recombinant strains towards simultaneous chemical production and CO2 assimilation in E. coli are limited and reported studies are questioned with critically low efficiency. In this context, we first expressed the ribulose-1,5-bisphosphate carboxylase / oxygenases (RuBisCOs) and phosphoribulokinases (PRKs) from the Calvin cycle to determine the ability to assimilate CO2 in E. coli with a value of -4.9 g-CO2 / g. to optimize -DCW. In order to improve the ability to assimilate CO2 and to expand the applicability of E. coli equipped with RuBisCO, we carried out a step-by-step integration of the recombinant genes (RuBisCO and PRK) and the introduction of the CRISPRi system to redirect the carbon flow to the RuBisCO path . The last strain, SSCI with CRISPRi targeted at zwf and pfkAB, improved the CO2 assimilation ability to -1.58 g-CO2 / g-DCW. Finally, E. coli equipped with RuBisCO were used for the simultaneous recycling of CO2 and the production of the high-quality molecules from various proteins, including 5-aminoleuvic acid (ALA) by ALA synthetase, carbon dioxide sequestration via carbonic anhydrase (CA) and 1,5 Diaminopentane (DAP) by lysine decarboxylase. This RuBisCO-equipped genomic E. coli neutralizer is nominated as a GREEN workhorse and offers great opportunities to be a microbial cell factory with a low carbon footprint.