Formate can be envisioned at the core of a carbon-neutral bioeconomy, where it is produced from CO2by (electro-)chemical means and converted into value-added products by enzymatic cascades or engineered microbes. A key step in expanding synthetic formate assimilation is its thermodynamically challenging reduction to formaldehyde. Here, we develop a two-enzyme route in which formate is activated to formyl phosphate and subsequently reduced to formaldehyde. Exploiting the promiscuity of acetate kinase andN-acetyl-γ-glutamyl phosphate reductase, we demonstrate this phosphate (Pi)-based route in vitro and in vivo. We further engineer a formyl phosphate reductase variant with improved formyl phosphate conversion in vivo by suppressing cross-talk with native metabolism and interface the Piroute with a recently developed formaldehyde assimilation pathway to enable C2 compound formation from formate as the sole carbon source inEscherichia coli. The Piroute therefore offers a potent tool in expanding the landscape of synthetic formate assimilation.