Yu Tonghuan, Takahashi Ukyo, Iwahashi Hitoshi
Key Laboratory of Yellow River and Huai River Water Environment and Pollution Control, Ministry of Education, Henan Key Laboratory for Environmental Pollution Control, International Joint Laboratory on Key Techniques in Water Treatment, School of Environment, Henan Normal University, Xinxiang 453007, China.
The United Graduate School of Agricultural Science, Gifu University, Yanagido 1-1, Gifu 501-1193, Japan.
J Fungi (Basel). 2022 Sep 27;8(10):1011. doi: 10.3390/jof8101011.
High-pressure carbon dioxide (HPCD), a novel non-thermal pasteurization technology, has attracted the attention of scientists due to its high pasteurization efficiency at a lower temperature and pressure. However, the inactivation mechanism has not been well researched, and this has hindered its commercial application. In this work, we used a sub-lethal HPCD condition (4.0 MPa, 30 °C) and a recovery condition (30 °C) to repair the damaged cells. Transcriptome analysis was performed by using RNA sequencing and gene ontology analysis to investigate the detailed lethal mechanism caused by HPCD treatment. RT-qPCR analysis was conducted for certain upregulated genes, and the influence of HPCD on protoplasts and single-gene deletion strains was investigated. Six major categories of upregulated genes were identified, including genes associated with the pentose phosphate pathway (oxidative phase), cell wall organization or biogenesis, glutathione metabolism, protein refolding, phosphatidylcholine biosynthesis, and AdoMet synthesis, which are all considered to be associated with cell death induced by HPCD. The inactivation or structure alteration of YNL194Cp in the organelle membrane is considered the critical reason for cell death. We believe this work contributes to elucidating the cell-death mechanism and providing a direction for further research on non-thermal HPCD sterilization technology.
高压二氧化碳(HPCD)是一种新型的非热杀菌技术,因其在较低温度和压力下具有较高的杀菌效率而受到科学家们的关注。然而,其失活机制尚未得到充分研究,这阻碍了其商业应用。在这项工作中,我们使用亚致死HPCD条件(4.0 MPa,30°C)和恢复条件(30°C)来修复受损细胞。通过RNA测序和基因本体分析进行转录组分析,以研究HPCD处理导致的详细致死机制。对某些上调基因进行了RT-qPCR分析,并研究了HPCD对原生质体和单基因缺失菌株的影响。确定了六类上调基因,包括与磷酸戊糖途径(氧化阶段)、细胞壁组织或生物合成、谷胱甘肽代谢、蛋白质重折叠、磷脂酰胆碱生物合成和腺苷甲硫氨酸合成相关的基因,这些基因都被认为与HPCD诱导的细胞死亡有关。细胞器膜中YNL194Cp的失活或结构改变被认为是细胞死亡的关键原因。我们相信这项工作有助于阐明细胞死亡机制,并为进一步研究非热HPCD杀菌技术提供方向。