The effects of chemical composition and pretreatment on Mg–Al hydrotalcites and alumina-supported MgO were evaluated for the gas-phase, self-condensation reaction of C3–C5 biomass-derived methyl ketones. We show that the selectivity toward the acyclic dimer enone and the cyclic enone trimer can be tuned by controlling the temperature of hydrotalcite calcination. Methyl ketone cyclization is promoted by Lewis acidic sites present on the hydrotalcite catalysts. XRD and thermal decomposition analysis reveal that the formation of periclase MgO starts above 623 K accompanied by complete disappearance of the hydrotalcite structure and is accompanied by an increase in hydroxyl condensation as the formation of well-crystallized periclase. 27Al MQMAS and 25Mg MAS NMR show that at progressively higher temperatures, Al3+ cations diffuses out of the octahedral brucite layers and incorporate into the tetrahedral and octahedral sites of the MgO matrix thereby creating defects to compensate the excess positive charge generated. The oxygen anions adjacent to the Mg2+/Al3+ defects become coordinatively unsaturated, leading to the formation of new basic sites. A kinetic isotope effect, kH/kD = 0.96, is observed at 473 K for the reaction of (CH3)2CO versus (CD3)2CO, which suggests that carbon–carbon bond formation leading to the dimer aldol product is the rate-determining step in the condensation reaction of methyl ketones. We also show that acid–base catalysts having similar reactivity and higher hydrothermal stability to that of calcined hydrotalcites can be achieved by creating defects in MgO crystallites supported alumina as a consequence of the diffusion of Al3+ cations into MgO. The physical properties of these materials are shown to be very similar to those of hydrotalcite calcined at 823 K.