Key Laboratory of Industrial Biotechnology, Ministry of Education and School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu 214122, China.
Appl Microbiol Biotechnol. 2013 Jul;97(14):6467-75. doi: 10.1007/s00253-013-4931-9. Epub 2013 May 5.
In the present study, we investigated the mode of cell response induced by D-limonene in Saccharomyces cerevisiae. D-limonene treatment was found to be accompanied by intracellular accumulation of reactive oxygen species (ROS). Since ROS impair cell membranes, an engineered strain with enhanced membrane biosynthesis exhibited a higher tolerance to D-limonene. Subsequent addition of an ROS scavenger significantly reduced the ROS level and alleviated cell growth inhibition. Thus, D-limonene-induced ROS accumulation plays an important role in cell death in S. cerevisiae. In D-limonene-treated S. cerevisiae strains, higher levels of antioxidants, antioxidant enzymes, and nicotinamide adenine dinucleotide phosphate (NADPH) were synthesized. Quantitative real-time PCR results also verified that D-limonene treatment triggered upregulation of genes involved in the antioxidant system and the regeneration of NADPH at the transcription level in S. cerevisiae. These data indicate that D-limonene treatment results in intracellular ROS accumulation, an important factor in cell death, and several antioxidant mechanisms in S. cerevisiae were enhanced in response to D-limonene treatment.
在本研究中,我们研究了 D-柠檬烯在酿酒酵母中诱导细胞反应的方式。研究发现,D-柠檬烯处理伴随着活性氧(ROS)的细胞内积累。由于 ROS 会损害细胞膜,因此具有增强膜生物合成的工程菌株对 D-柠檬烯表现出更高的耐受性。随后添加 ROS 清除剂可显著降低 ROS 水平并减轻细胞生长抑制。因此,D-柠檬烯诱导的 ROS 积累在酿酒酵母细胞死亡中起重要作用。在 D-柠檬烯处理的酿酒酵母菌株中,合成了更高水平的抗氧化剂、抗氧化酶和烟酰胺腺嘌呤二核苷酸磷酸(NADPH)。定量实时 PCR 结果还证实,D-柠檬烯处理在转录水平上触发了与抗氧化系统和 NADPH 再生相关的基因的上调。这些数据表明,D-柠檬烯处理导致细胞内 ROS 积累,这是细胞死亡的一个重要因素,并且 D-柠檬烯处理增强了酿酒酵母中的几种抗氧化机制。