为探究酵母在氧化胁迫下外源添加原花色素对胞内海藻糖积累以及cAMP-PKA信号转导途径的影响。在氧化胁迫条件下,外加添加1g/L的原花色素,分别分析胞内海藻糖积累量、上游信号转导关键元件PKA,腺苷酸环化酶的转录规律及cAMP的水平。添加H2O2后再添加1 g/L的原花色素的发酵体系中酵母细胞内海藻糖产量是空白对照组的3倍,cAMP产量和腺苷酸环化酶的酶活是空白对照组的2.5倍,CYR1和BCY1基因的相对表达量较空白组均呈现明显升高趋势。cAMP会与PKA调节的亚基BCY1相结合,释放出催化亚基从而激活PKA,产生海藻糖保护细胞抵抗不利环境。该研究为以后深入探究氧化胁迫下原花色素对酿酒酵母的保护机制奠定基础,也为以后探究酿酒酵母在面临其他胁迫条件下如何抵御外界环境变化奠定基础。
The effects of proanthocyanins on intracellular trehalose accumulation and cAMP signal transduction pathway in Saccharomyces cerevisiae were investigated under oxidative stress. The trehalose accumulation amount, upstream signal transduction key element (PKA), transcription rule of adenylate cyclase and cAMP level were analyzed. With addition of 0.5 mmol/L H2O2 and 1 g/L proanthocyanidin, the amount of intracellular trehalose in S. cerevisiae increased by 2 times, cAMP concentration and adenylate cyclase activity by 1.5 times along with a significant increase of the relative expression levels of CYR1 and BCY1. It was found that cAMP bound to PKA-regulated subunit BCY1, led to release of the catalytic subunit that activated PKA, followed by trehalose accumulation protecting the cell against adverse environments. This study provides a base for further investigation of S. cerevisiae protection by proanthocyanins against oxidative and other stresses.
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