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通过异源表达汉逊德巴利酵母的过氧化氢酶A和T提高出芽酵母对氧化应激的耐受性

Tolerance to Oxidative Stress in Budding Yeast by Heterologous Expression of Catalases A and T from Debaryomyces hansenii.

作者信息

González James, Castillo Román, García-Campos Miguel Angel, Noriega-Samaniego Diego, Escobar-Sánchez Viviana, Romero-Aguilar Lucero, Alba-Lois Luisa, Segal-Kischinevzky Claudia

机构信息

Departamento de Biología Celular, Facultad de Ciencias, Universidad Nacional Autónoma de México. Avenida Universidad 3000, Cd. Universitaria, 04510, Coyoacán, Ciudad de México, México.

Departamento de Bioquímica, Facultad de Medicina, Facultad de Medicina, Universidad Nacional Autónoma de México. Avenida Universidad 3000, Cd. Universitaria, 04510, Coyoacán, Ciudad de México, México.

出版信息

Curr Microbiol. 2020 Dec;77(12):4000-4015. doi: 10.1007/s00284-020-02237-3. Epub 2020 Oct 16.

Abstract

The function of catalases A and T from the budding yeast Saccharomyces cerevisiae (ScCta1 and ScCtt1) is to decompose hydrogen peroxide (HO) to mitigate oxidative stress. Catalase orthologs are widely found in yeast, suggesting that scavenging HO is crucial to avoid the oxidative damage caused by reactive oxygen species (ROS). However, the function of catalase orthologs has not yet been experimentally characterized in vivo. Here, we heterologously expressed Debaryomyces hansenii DhCTA1 and DhCTT1 genes, encoding ScCta1 and ScCtt1 orthologs, respectively, in a S. cerevisiae acatalasemic strain (cta1Δ ctt1Δ). We performed a physiological analysis evaluating growth, catalase activity, and HO tolerance of the strains grown with glucose or ethanol as carbon source, as well as under NaCl stress. We found that both genes complement the catalase function in S. cerevisiae. Particularly, the strain harboring DhCTT1 showed improved growth when ethanol was used as carbon source both in the absence or presence of salt stress. This phenotype is attributed to the high catalase activity of DhCtt1 detected at the exponential growth phase, which prevents intracellular ROS accumulation and confers oxidative stress resistance.

摘要

来自出芽酵母酿酒酵母(ScCta1和ScCtt1)的过氧化氢酶A和T的功能是分解过氧化氢(HO)以减轻氧化应激。过氧化氢酶直系同源物在酵母中广泛存在,这表明清除HO对于避免活性氧(ROS)引起的氧化损伤至关重要。然而,过氧化氢酶直系同源物的功能尚未在体内进行实验表征。在这里,我们在酿酒酵母无过氧化氢酶菌株(cta1Δ ctt1Δ)中异源表达了分别编码ScCta1和ScCtt1直系同源物的汉逊德巴利酵母DhCTA1和DhCTT1基因。我们进行了一项生理分析,评估了以葡萄糖或乙醇作为碳源生长的菌株以及在NaCl胁迫下的生长、过氧化氢酶活性和HO耐受性。我们发现这两个基因都能在酿酒酵母中补充过氧化氢酶功能。特别是,携带DhCTT1的菌株在有无盐胁迫的情况下,以乙醇作为碳源时均表现出改善的生长。这种表型归因于在指数生长期检测到的DhCtt1的高过氧化氢酶活性,它可防止细胞内ROS积累并赋予氧化应激抗性。

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