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Biomarkers for detecting nitrogen deficiency during alcoholic fermentation in different commercial wine yeast strains

机译：用于检测不同商业葡萄酒酵母菌株酒精发酵过程中氮缺乏的生物标志物

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摘要

Nitrogen deficiencies in grape musts are one of the main causes of stuck or sluggish wine fermentations. Several putative biomarkers were tested in order to analyze their appropriateness to detect nitrogen stress in the yeast. To this aim, four commercial wine strains (PDM, ARM, RVA and TTA) were grown in a synthetic grape must with different nitrogen concentrations. Trehalose accumulation, arginase activity and the expression of eleven genes were tested in these wine strains, known to have different nitrogen requirements. The overall response of the four strains was similar, with differences in response intensity (PDM and RVA with higher intensity) and response time (which was also related with nitrogen consumption time). Trehalose response was mostly related to entry into the stationary phase, whereas arginase activity was responsive to nitrogen depletion, although its measurement is too complicated to be used for routine monitoring during winemaking. The expression of the genes DAL4, DAL5, DUR3 and GAP1 was clearly related to nitrogen depletion and thus, GAP1 and DAL4 were selected as markers of nitrogen deficiency. In order to adapt expression analysis to winemaking conditions, the original strains were transformed into reporter strains based on the expression of green fluorescent protein (GFP) under control of the promoters for CAP1 and DAL4. The transformants had a similar fermentative capacity to the parental strains and were able to detect alterations in yeast physiological status due to nitrogen limitations.
机译：葡萄汁中的氮缺乏是葡萄酒发酵停滞或缓慢的主要原因之一。测试了几种推定的生物标记，以分析其检测酵母中氮胁迫的适当性。为此，在具有不同氮浓度的合成葡萄汁中种植了四种商业葡萄酒菌株（PDM，ARM，RVA和TTA）。在这些已知氮需求不同的葡萄酒菌株中测试了海藻糖的积累，精氨酸酶活性和11个基因的表达。四种菌株的总体响应相似，但响应强度（PDM和RVA强度更高）和响应时间（也与氮消耗时间有关）有所不同。海藻糖反应主要与进入固定相有关，而精氨酸酶活性则对氮耗竭有反应，尽管其测量太复杂，无法用于酿酒过程中的常规监测。 DAL4，DAL5，DUR3和GAP1基因的表达与氮耗竭明显相关，因此，选择GAP1和DAL4作为氮缺乏的标志。为了使表达分析适应酿酒条件，在CAP1和DAL4的启动子的控制下，基于绿色荧光蛋白（GFP）的表达，将原始菌株转化为报告菌株。转化子具有与亲本菌株相似的发酵能力，并且能够检测由于氮限制而引起的酵母生理状态的改变。

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  • 来源
    《Food microbiology》 |2013年第1期|227-237|共11页
  • 作者

    Alicia Gutierrez; Rosana Chiva; Gemma Beltran; Albert Mas; Jose Manuel Guillamon;

  • 作者单位

    Departamento de Biotecnologia de los alimentos, Instituto de Agroquimica y Tecnologia de los Alimentos (CSIC), Avda. Agustin Escardino 7, E-46980 Paterna,Valencia, Spain;

    Departamento de Biotecnologia de los alimentos, Instituto de Agroquimica y Tecnologia de los Alimentos (CSIC), Avda. Agustin Escardino 7, E-46980 Paterna,Valencia, Spain;

    Biotecnologia Enologica, Departament de Bioquimica i Biotecnologia, Facultat d'Enologia, Universitat Rovira i Virgili, Marcel-li Domingo s, 43007 Tarragona, Spain;

    Biotecnologia Enologica, Departament de Bioquimica i Biotecnologia, Facultat d'Enologia, Universitat Rovira i Virgili, Marcel-li Domingo s, 43007 Tarragona, Spain;

    Departamento de Biotecnologia de los alimentos, Instituto de Agroquimica y Tecnologia de los Alimentos (CSIC), Avda. Agustin Escardino 7, E-46980 Paterna,Valencia, Spain;

  • 收录信息 美国《科学引文索引》(SCI);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    nitrogen starvation; arginase activity; gene expression; general amino acid permease; gfp-biosensor; fluorescence reporter genes; yeast assimilable nitrogen;

    机译：氮饥饿精氨酸酶活性;基因表达;一般氨基酸通透酶;gfp生物传感器;荧光报告基因;酵母同化氮;

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