Departamento de Biotecnología, Instituto de Agroquímica y Tecnología de Alimentos, CSIC, Apartado de Correos, 73. Burjassot (Valencia). E-46100, Spain.
Microb Cell Fact. 2012 Jan 9;11:4. doi: 10.1186/1475-2859-11-4.
In the yeast biomass production process, protein carbonylation has severe adverse effects since it diminishes biomass yield and profitability of industrial production plants. However, this significant detriment of yeast performance can be alleviated by increasing thioredoxins levels. Thioredoxins are important antioxidant defenses implicated in many functions in cells, and their primordial functions include scavenging of reactive oxygen species that produce dramatic and irreversible alterations such as protein carbonylation.
In this work we have found several proteins specifically protected by yeast Thioredoxin 2 (Trx2p). Bidimensional electrophoresis and carbonylated protein identification from TRX-deficient and TRX-overexpressing cells revealed that glycolysis and fermentation-related proteins are specific targets of Trx2p protection. Indeed, the TRX2 overexpressing strain presented increased activity of the central carbon metabolism enzymes. Interestingly, Trx2p specifically preserved alcohol dehydrogenase I (Adh1p) from carbonylation, decreased oligomer aggregates and increased its enzymatic activity.
The identified proteins suggest that the fermentative capacity detriment observed under industrial conditions in T73 wine commercial strain results from the oxidative carbonylation of specific glycolytic and fermentation enzymes. Indeed, increased thioredoxin levels enhance the performance of key fermentation enzymes such as Adh1p, which consequently increases fermentative capacity.
在酵母生物量生产过程中,蛋白质羰基化作用会产生严重的负面影响,因为它会降低生物量产量和工业生产工厂的盈利能力。然而,通过增加硫氧还蛋白的水平可以减轻酵母性能的这种显著损害。硫氧还蛋白是一种重要的抗氧化防御物质,涉及细胞中的许多功能,其原始功能包括清除产生剧烈和不可逆改变的活性氧物质,如蛋白质羰基化。
在这项工作中,我们发现了几种酵母硫氧还蛋白 2(Trx2p)特异性保护的蛋白质。二维电泳和 TRX 缺陷和 TRX 过表达细胞中的羰基化蛋白鉴定表明,糖酵解和发酵相关蛋白是 Trx2p 保护的特定靶标。事实上,TRX2 过表达菌株表现出中央碳代谢酶活性增加。有趣的是,Trx2p 特异性地保护了醇脱氢酶 I(Adh1p)免受羰基化,减少了寡聚体聚集并增加了其酶活性。
所鉴定的蛋白质表明,在 T73 葡萄酒商业菌株的工业条件下观察到的发酵能力下降是由于特定糖酵解和发酵酶的氧化羰基化。事实上,增加硫氧还蛋白水平可以增强关键发酵酶如 Adh1p 的性能,从而提高发酵能力。