biostudies-literatureDinh HCore Research for Evolutional Science and TechnologyJapan Society for the Promotion of Science9088-9100https://www.ebi.ac.uk/biostudies/studies/S-EPMC8161546biostudies-literatureUnknown11(34)In contrast to the dilute conditions employed for <i>in vitro</i> biochemical studies, enzymes are spatially organized at high density in cellular micro-compartments. In spite of being crucial for cellular functions, enzymatic reactions in such highly packed states have not been fully addressed. Here, we applied a protein adaptor to assemble a single type of monomeric enzyme on a DNA scaffold in the packed or dispersed states for carbonic anhydrase. The enzymatic reactions proceeded faster in the packed than in the dispersed state. Acceleration of the reaction in the packed assembly was more prominent for substrates with higher hydrophobicity. In addition, carbonic anhydrase is more tolerant of inhibitors in the packed assembly. Such an acceleration of the reaction in the packed state overChemical scienceEnhanced enzymatic activity exerted by a packed assembly of a single type of enzyme.PMC8161546JPMJCR18H519H0465317H01213Nakata EDinh HKinoshita MMorii TMutsuda-Zapater KSaimura MfalseEnhanced enzymatic activity exerted by a packed assembly of a single type of enzyme.In contrast to the dilute conditions employed for <i>in vitro</i> biochemical studies, enzymes are spatially organized at high density in cellular micro-compartments. In spite of being crucial for cellular functions, enzymatic reactions in such highly packed states have not been fully addressed. Here, we applied a protein adaptor to assemble a single type of monomeric enzyme on a DNA scaffold in the packed or dispersed states for carbonic anhydrase. The enzymatic reactions proceeded faster in the packed than in the dispersed state. Acceleration of the reaction in the packed assembly was more prominent for substrates with higher hydrophobicity. In addition, carbonic anhydrase is more tolerant of inhibitors in the packed assembly. Such an acceleration of the reaction in the packed state over2020-01-01T00:00:00Z2020 Jul2025-04-18T23:00:08.857Z2022-02-10T13:47:34.821ZS-EPMC81615463409419010.1039/d0sc03498c