Abstract Rhamnulose‐1‐phosphate aldolases (RhuA) were the first dihydroxyacetone phosphate (DHAP) aldolases to be demonstrated to efficiently accept a ketone as the electrophile, providing the highly sought‐after chiral tertiary alcohol motif. It has also been found that this enzyme family can use dioxygen as an electrophile, forcing to work under inert atmosphere conditions when the target electrophile is poorly reactive. However, these features have not been explored in all DHAP‐aldolase families. Here, we present a study of E. coli K12 fuculose‐1‐phosphate‐ (FucA K12 ), fructose‐1,6‐ bis phosphate‐ (FruA K12 ) and tagatose‐1,6‐bisphosphate (TagA K12 ) aldolases. For the latter two enzymes, this was made possible by the development of a triose phosphate isomerase (TPI)‐deleted E coli strain, which prevents DHAP isomerization into d ‐glyceraldehyde 3‐phosphate (G3P), its natural electrophile. Like RhuA, TagA K12 was found to be able to oxidize DHAP into hydroxypyruvaldehyde phosphate under dioxygen, leading to the corresponding diulose. On the other hand, FucA K12 was revealed to be capable of converting ketone electrophiles with greater stereoselectivity than RhuA, and without the requirement of an inert atmosphere, since it was unable to oxidize DHAP. Finally, FruA K12 proved unable either to oxidize DHAP and or to accept ketones as electrophiles.