Simão Neto, FranciscoSousa Junior, Paulo Gonçalves deda Silva Filho, Carlos José AlvesPinheiro Coutinho, LucasMelo, Rafael Leandro FernandesRocha Martín, JavierRios, Maria Alexsandra de SousaSanders Lopes, Ada AméliaMonteiro, Norberto de K. V.de Mattos, Marcos CarlosSerafim, Leonardo FariasSousa dos Santos, José Cleiton2025-03-252025-03-252024-04-01Simão Neto, F., Sousa Junior, P. G. d., da Silva Filho, C. J. A., Pinheiro Coutinho, L., Melo, R. L. F., Rocha-Martin, J., Rios, M. A. d. S., Sanders Lopes, A. A., Monteiro, N. d. K. V., de Mattos, M. C., Serafim, L. F., & Sousa dos Santos, J. C. (2023). Green Enzymatic Synthesis of Geranyl Butyrate: Process Optimization and Mechanistic Insights. ACS Omega. https://doi.org/10.1021/ACSOMEGA.3C084832470-134310.1021/acsomega.3c08483https://hdl.handle.net/20.500.14352/118927We gratefully acknowledge the following Brazilian Agencies for Scientific and Technological Development: Fundação Cearense de Apoio ao Desenvolvimento Científico e Tecnológico (FUNCAP) (PS1-0186-00216.01.00/21; UNI-0210-00537.01.00/23), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) (311062/2019-9; 440891/2020-5; 307454/2022-3), and Coordenação de Aperfeiçoamento de Ensino Superior (CAPES) (finance code 001). The authors thank the Northeastern Center for Application and Use of NMR (CENAUREMN) for NMR spectroscopy.Flavor esters are organic compounds widely used in the food industry to enhance the aroma and taste of products. However, most chemical processes for the production of these flavoring compounds use toxic organic solvents. Some organic solvents derived from petroleum can leave behind residual traces in food products, which may raise concerns about potential health risks and contamination. In this study, we employ Eversa Transform 2.0, a commercial lipase derived from the lipase from Thermomyces lanuginosus, to produce geranyl butyrate in aqueous media. The chemical process was optimized using the Taguchi method, and a conversion of 93% was obtained at the optimal reaction conditions of: 1:5 molar ratio (v/v), 15% biocatalyst load (w/w), at 50 °C, in 6 h. Classic (molecular dynamics) and quantum (density functional theory) simulations unveiled amino acid residues involved in the stabilization of the enzyme–substrate complex. Detailed QM/MM mechanistic studies identified the nucleophilic attack of the deacylation reaction as the rate-limiting step of the entire mechanism, which has a free energy barrier of 14.0 kcal/mol.engAttribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/journal articlehttps://doi.org/10.1021/acsomega.3c08483https://pubs.acs.org/doi/10.1021/acsomega.3c08483open access612.393613.2641.1577.15547663/665Chemical reactionsFlavorOrganic compoundsOrganic reactionsPeptides and proteinsAlimentaciónBioquímica (Biología)Tecnología de los alimentosQuímica orgánica (Química)3309 Tecnología de Los Alimentos2302.90 Bioquímica de Alimentos2302.09 Enzimología2306 Química Orgánica3309.10 Aroma y Sabor