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Chitosan-Based Nanoencapsulation of Ocimum americanum Essential Oil as Safe Green Preservative Against Fungi Infesting Stored Millets, Aflatoxin B1 Contamination, and Lipid Peroxidation

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Abstract

Present study deals with the first-time report on encapsulation of Ocimum americanum essential oil (OAEO) into chitosan matrix with enhanced antifungal, aflatoxin B1 (AFB1) inhibition, antioxidant activity, and in situ efficacy in the millet food system. GC–MS analysis suggested citral (66.72%) as the major component of OAEO. Physicochemical characterizations through SEM, FTIR, and XRD analyses confirmed the successful loading of OAEO into chitosan nanoemulsion (OAEO-CsNe). In vitro release profile of nanoencapsulated OAEO exhibited biphasic burst and controlled volatilisation, a prerequisite for long-term antifungal effect in the stored food system. OAEO-CsNe completely inhibited the growth and AFB1 production of Aspergillus flavus at 0.2 and 0.175 μL/mL, respectively. Inhibition of ergosterol biosynthesis followed by the release of vital cellular ions, and 260, 280 nm absorbing materials from AFLHPSi-1 cells suggested plasma membrane as a potential site of antifungal action of OAEO-CsNe. Significant reduction of cellular methylglyoxal (an AFB1 inducer) level in AFLHPSi-1 cells after fumigation with OAEO-CsNe confirmed the novel biochemical mechanism of anti-aflatoxigenic activity. Additionally, in silico modelling of citral (major component of OAEO) with Ver-1 and Omt-A proteins suggested the hydrogen bond-dependent molecular interaction for inhibition of AFB1 biosynthesis. OAEO-CsNe showed significant in situ antifungal, anti-aflatoxigenic, and lipid peroxidation-suppressing potentialities without altering the organoleptic and germination properties of Setaria italica seeds. Moreover, the appreciative safety profile (LD50 = 11,162.06 μL/kg) of OAEO-CsNe in a mammalian model system (Mus musculus) strengthens its recommendation as an effective green preservative against fungal infestation, AFB1 contamination, and reactive oxygen species-mediated lipid peroxidation in stored food commodities.

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Acknowledgements

Bijendra Kumar Singh is grateful to Head, Department of Botany, and Principal of Feroze Gandhi College, Raebareli, Uttar Pradesh, India for kind support. Authors are thankful to Head and Coordinator, CAS, Department of Botany, ISLS, for providing laboratory facilities, and Central Instrument Facility, Indian Institute of Technology, Banaras Hindu University, Varanasi for SEM, FTIR and XRD analysis.

Funding

Bijendra Kumar Singh is thankful to the Council of Scientific and Industrial Research (CSIR) (grant no.: 09/013(0920)/2019-EMR-I), New Delhi, India for providing research fellowship.

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Authors and Affiliations

  1. Laboratory of Herbal Pesticides, Centre of Advanced Study (CAS) in Botany, Institute of Science, Banaras Hindu University, Varanasi, 221005, India

    Bijendra Kumar Singh, Shikha Tiwari, Akash Maurya & Nawal Kishore Dubey

  2. Department of Botany, Burdwan Raj College, Purba Bardhaman, West Bengal, Bardhaman, 713104, India

    Somenath Das

  3. Department of Botany, Feroze Gandhi College, Raebareli, 229001, India

    Bijendra Kumar Singh

  4. Department of Botany, K. S. Saket P. G. College, Ayodhya, 224123, India

    Vipin Kumar Singh

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Contributions

Bijendra Kumar Singh: conceptualization, methodology, writing original draft, data curation, and funding acquisition. Shikha Tiwari: data curation. Akash Maurya: data curation. Somenath Das: review and editing. Vipin Kumar Singh: review and editing. Nawal Kishore Dubey: supervision and writing—review and editing.

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Correspondence to Nawal Kishore Dubey.

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Singh, B.K., Tiwari, S., Maurya, A. et al. Chitosan-Based Nanoencapsulation of Ocimum americanum Essential Oil as Safe Green Preservative Against Fungi Infesting Stored Millets, Aflatoxin B1 Contamination, and Lipid Peroxidation. Food Bioprocess Technol 16, 1851–1872 (2023). https://doi.org/10.1007/s11947-023-03008-1

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