The Dual-Functional Monoterpenes D-Carvone and Carveol in Citrus-Drop Essential Oil Reprogram Listeria monocytogenes Bioﬁlms via an Escape-Induction and Metabolic-Lethality Synergistic Strategy

Jiajing Guo; Wenbin Xiao; Shenghua Ding; Yanjiao Fu; Yanfang Liao; Yunlong Ding; Ziyi Fan; Gaoyang Li; Donglin Su; Yang Shan; Siew Young Quek

doi:10.1016/j.eng.2026.05.022

Engineering ›› :202605022 DOI: 10.1016/j.eng.2026.05.022

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The Dual-Functional Monoterpenes D-Carvone and Carveol in Citrus-Drop Essential Oil Reprogram Listeria monocytogenes Bioﬁlms via an Escape-Induction and Metabolic-Lethality Synergistic Strategy

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Abstract

Listeria monocytogenes (LM) poses a signiﬁcant threat to food safety due to its ability to form persistent bioﬁlms. Thus, developing efﬁcient and green strategies to combat bioﬁlms is crucial in combating LM. This study evaluates the antibioﬁlm activity and mechanisms of essential oils (EOs) extracted from citrus physiological fruit drop (CPFD), an agricultural byproduct, against LM. Among eight CPFD-EOs, Ougan (Citrus reticulata cv. Suavissima) fruit drop essential oil (OGEO) exhibited the highest extraction yield (3.18%) and was found to have a unique chemical proﬁle characterized by enrichment in oxygenated monoterpenes, particularly D-carvone and carveol. OGEO demonstrated the strongest broad-spectrum antibacterial and antibioﬁlm activity, with two of its components, D-carvone and carveol, showing signif- icant synergistic antibacterial effects (the fractional inhibitory concentration (FIC) index = 0.5). More speciﬁcally, OGEO and its monoterpenes damaged bacterial cell membranes and disrupted bioﬁlm archi- tecture. Integrated transcriptomic and proteomic analyses revealed a multi-target mechanism: OGEO simultaneously inhibits carbon metabolism, nucleotide synthesis, adenosine triphosphate-binding cas- sette (ABC) transporters, and cell wall biosynthesis, while enhancing bacterial chemotaxis and ﬂagellar assembly, inducing bioﬁlm-embedded cells to ‘‘escape” from the protective matrix into a vulnerable planktonic state. D-Carvone possibly acts by disrupting cell wall integrity and inducing this ‘‘escape” (structural stripping + escape induction), while carveol mainly causes the collapse of glycolysis and amino acid metabolism (metabolic lethality). The synergy of these compounds constitutes a novel escape induction + lethal clearance synergistic model against bioﬁlms. Our study demonstrates that CPFD- derived EOs, particularly OGEO, are a promising source of natural antibioﬁlm agents. Their multi- target synergistic action effectively combats LM bioﬁlms, providing a theoretical basis for the valorization of agricultural waste and the development of novel food antimicrobial strategies.

Keywords

Listeria monocytogenes / Bioﬁlm / Citrus physiological fruit drop / D-Carvone / Carveol

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Jiajing Guo, Wenbin Xiao, Shenghua Ding, Yanjiao Fu, Yanfang Liao, Yunlong Ding, Ziyi Fan, Gaoyang Li, Donglin Su, Yang Shan, Siew Young Quek. The Dual-Functional Monoterpenes D-Carvone and Carveol in Citrus-Drop Essential Oil Reprogram Listeria monocytogenes Bioﬁlms via an Escape-Induction and Metabolic-Lethality Synergistic Strategy. Engineering 202605022 DOI:10.1016/j.eng.2026.05.022

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