Niu Liyuan, Nomura Kazuki, Iwahashi Hitoshi, Matsuoka Hiroyuki, Kawachi Satoshi, Suzuki Yoshihisa, Tamura Katsuhiro
United Graduate School of Agricultural Science, Gifu University, Gifu, Japan.
United Graduate School of Agricultural Science, Gifu University, Gifu, Japan.
Biophys Chem. 2017 Dec;231:79-86. doi: 10.1016/j.bpc.2017.03.003. Epub 2017 Mar 8.
Petit-High Pressure Carbon Dioxide (p-HPCD) is a promising nonthermal technology for foods pasteurization. Cluster analysis of gene expression profiles of Saccharomyces cerevisiae exposed to various stresses exhibited that gene expression profile for p-HPCD stress (0.5MPa, 25°C) was grouped into a cluster including profiles for Sodium Dodecyl Sulfate and Roundup herbicide. Both are detergents that can disorder membrane structurally and functionally, which suggests that cell membrane may be a target of p-HPCD stress to cause cell growth inhibition. Through metabolomic analysis, amount of S-Adenosylmethionine (AdoMet) that is used as methyl donor to participate in phosphatidylcholine synthesis via phosphatidylethanolamine (PE) methylation pathway, was increased after p-HPCD treatment for 2h. The key gene OPI3 encoding phospholipid methyltransferase that catalyzes the last two steps in PE methylation pathway was confirmed significantly induced by RT-PCR. Transcriptional expression of genes (MET13, MET16, MET10, MET17, MET6 and SAM2) related to AdoMet biosynthesis was also significantly induced. Choline as the PC precursor and ethanolamine as PE precursor in Kennedy pathway were also found increased under p-HPCD condition. We also found that amounts of most of amino acids involving protein synthesis were found decreased after p-HPCD treatment for 2h. Moreover, morphological changes on cell surface were observed by scanning electron microscope. In conclusion, the effects of p-HPCD stress on cell membrane appear to be a very likely cause of yeast growth inhibition and the enhancement of PC synthesis could contribute to maintain optimum structure and functions of cell membrane and improve cell resistance to inactivation.
微高压二氧化碳(p-HPCD)是一种很有前景的用于食品巴氏杀菌的非热技术。对暴露于各种应激条件下的酿酒酵母基因表达谱进行聚类分析表明,p-HPCD应激(0.5MPa,25°C)的基因表达谱被归为一个聚类,其中包括十二烷基硫酸钠和农达除草剂的基因表达谱。这两种都是能在结构和功能上扰乱细胞膜的去污剂,这表明细胞膜可能是p-HPCD应激导致细胞生长抑制的靶点。通过代谢组学分析,在p-HPCD处理2小时后,用作甲基供体通过磷脂酰乙醇胺(PE)甲基化途径参与磷脂酰胆碱合成的S-腺苷甲硫氨酸(AdoMet)的量增加。通过逆转录-聚合酶链反应(RT-PCR)证实,催化PE甲基化途径最后两步的磷脂甲基转移酶的关键基因OPI3被显著诱导。与AdoMet生物合成相关的基因(MET13、MET16、MET10、MET17、MET6和SAM2)的转录表达也被显著诱导。在p-HPCD条件下,还发现肯尼迪途径中作为PC前体的胆碱和作为PE前体的乙醇胺增加。我们还发现,在p-HPCD处理2小时后,参与蛋白质合成的大多数氨基酸的量减少。此外,通过扫描电子显微镜观察到细胞表面的形态变化。总之,p-HPCD应激对细胞膜的影响似乎很可能是酵母生长抑制的原因,而PC合成的增强可能有助于维持细胞膜的最佳结构和功能,并提高细胞的抗失活能力。