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The Impact of D-limonene on Cell Membrane Barrier of Pichia kluyveri Y-11519 from Sichuan Pickles

Chaoyi Zeng, Atittaya Tandhanskul, Samatcha Krungkaew, Tanawan Likhanapaiboon, Witthawat Kasayapanan, Watanya Chaisayan, Patchanee Yasurin, Jie Tang, Theerawut Phusantisampan, Atthasit Tawai


Sichuan pickles (SCP) are a traditional method of preserving vegetables in China. Through the spontaneous fermentation of microorganisms in brine water, it forms a unique flavor to meet with food industry’s requirements. However, the microorganisms in salt water determine the quality of SCP, and the film-forming phenomenon is considered to be the key to the spoilage of SCP, which seriously restricts the industrial development of SCP. We have noticed that in folk, lemon peel is often added to pickles to prevent the appearance of the film. Currently, the extract D-limonene from orange or lemon peel is recognized as a Generally Recognized as Safe (GRAS) food additive and exhibits broad-spectrum antimicrobial properties. However, there have been no reports on the effects of D-limonene on Pichia kluyveri (P. kluyveri), the microorganism responsible for the "film-forming" phenomenon in SCP. In this study, D-limonene was used to treat P. kluyveri Y-11519, a membranous microorganism of SCP, and the cell morphology, surface charge, membrane potential, and intracellular macromolecule leakage before and after treatment were observed. The results showed that the minimum inhibitory concentration of D-limonene against P. kluyveri Y-11519 was 20 μL/mL, and the minimum fungicidal concentration was 40 μL/mL. After treatment with this concentration of D-limonene, the growth of P. kluyveri Y-11519 cells was delayed, cells exhibited deformation and shrinkage, cell membrane integrity was compromised, permeability increased, intracellular substances leaked, ultimately leading to cell death.


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DOI: 10.14416/j.asep.2024.06.014


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