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低温发酵条件下葡萄酒酵母适应功能分析及相关基因鉴定

Functional analysis to identify genes in wine yeast adaptation to low-temperature fermentation.

机构信息

Biotecnologia Enològica, Departament de Bioquímica i Biotecnologia, Facultat de Enologia, Universitat Rovira i Virgili, Tarragona, Spain.

出版信息

J Appl Microbiol. 2012 Jul;113(1):76-88. doi: 10.1111/j.1365-2672.2012.05308.x. Epub 2012 May 10.

Abstract

AIMS

To identify genes and proteins involved in adaptation to low-temperature fermentations in a commercial wine yeast.

METHODS AND RESULTS

Nine proteins were identified as representing the most significant changes in proteomic maps during the first 24 h of fermentation at low (13°C) and standard temperature (25°C). These proteins were mainly involved in stress response and in glucose and nitrogen metabolism. Transcription analysis of the genes encoding most of these proteins within the same time frame of wine fermentation presented a good correlation with proteomic data. Knockout and overexpressing strains of some of these genes were constructed and tested to evaluate their ability to start the fermentation process. The strain overexpressing ILV5 improved its fermentation activity in the first hours of fermentation. This strain showed a quicker process of mitochondrial degeneration, an altered intracellular amino acid profile and laxer nitrogen catabolite repression regulation.

CONCLUSIONS

The proteomic and transcriptomic analysis is useful to detect key molecular adaptation mechanisms of biotechnological interest for industrial processes. ILV5 gene seems to be important in wine yeast adaptation to low-temperature fermentation.

SIGNIFICANCE AND IMPACT OF THE STUDY

This study provides information that might help improve the future performance of wine yeast, either by genetic modification or by adaptation during industrial production.

摘要

目的

鉴定参与商业葡萄酒酵母低温发酵适应的基因和蛋白质。

方法和结果

在低温(13°C)和标准温度(25°C)下发酵的前 24 小时内,蛋白质组图谱中鉴定出 9 种蛋白质发生了最显著的变化。这些蛋白质主要参与应激反应以及葡萄糖和氮代谢。在相同的葡萄酒发酵时间框架内,对编码这些蛋白质的基因进行转录分析,与蛋白质组数据有很好的相关性。构建并测试了这些基因的一些敲除和过表达菌株,以评估它们启动发酵过程的能力。过表达 ILV5 的菌株提高了其在发酵初期的发酵活性。该菌株表现出更快的线粒体退化过程、改变的细胞内氨基酸谱和更宽松的氮分解代谢物抑制调控。

结论

蛋白质组学和转录组学分析可用于检测对工业过程具有生物技术意义的关键分子适应机制。ILV5 基因似乎在葡萄酒酵母对低温发酵的适应中很重要。

研究的意义和影响

本研究提供的信息可能有助于提高未来葡萄酒酵母的性能,无论是通过遗传修饰还是在工业生产过程中的适应。

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