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冰酒发酵过程中酿酒酵母醛脱氢酶对乙醛胁迫的响应

Response of wine yeast (Saccharomyces cerevisiae) aldehyde dehydrogenases to acetaldehyde stress during Icewine fermentation.

作者信息

Pigeau G M, Inglis D L

机构信息

Department of Biological Sciences, Brock University, St. Catharines, Ontario, Canada.

出版信息

J Appl Microbiol. 2007 Nov;103(5):1576-86. doi: 10.1111/j.1365-2672.2007.03381.x.

Abstract

AIMS

We previously reported that the aldehyde dehydrogenase encoded by ALD3 but not ALD6 was responsible, in part, for the increased acetic acid found in Icewines based on the expression profile of these genes during fermentation. We have now completed the expression profile of the remaining yeast aldehyde dehydrogenase genes ALD2, ALD4 and ALD5 during these fermentations to determine their contribution to acetic acid production. The contribution of acetaldehyde stress as a signal to stimulate ALD expression during these fermentations was investigated for all ALD genes. The expression of glycerol-3-phosphate encoded by GPD2 was also followed during these fermentations to determine its role in addition to the role we already identified for GPD1 in the elevated glycerol produced during Icewine fermentation.

METHODS AND RESULTS

Icewine juice (38.5 degrees Brix, 398 +/- 5 g l(-1) sugar), diluted Icewine juice (20.8 degrees Brix, 196 +/- 4 g l(-1) sugar) and the diluted juice with sugar levels equal to the original Icewine juice (36.6 degrees Brix, 395 +/- 6 g l(-1) sugar) were fermented in duplicate using the commercial wine yeast K1-V1116. Acetic acid and glycerol production increased 8.4- and 2.7-fold in the Icewine vs the diluted juice fermentation, respectively, accompanied by a fourfold transient increase in acetaldehyde in the Icewine condition during the first week. Both mitochondrial aldehyde dehydrogenases encoded by ALD4 and ALD5 were expressed, with ALD5 expression highest at the start of all fermentations and ALD4 expression increasing during the first week of each condition. ALD2, ALD4, ALD5 and GPD2 showed no differential expression between the three fermentation conditions indicating their lack of involvement in elevating acetic acid and glycerol in Icewine. When yeast fermenting the diluted fermentation was exposed to exogenous acetaldehyde, the transient spike in acetaldehyde increased the expression of ALD3 but this response alone was not sufficient to cause an increase in acetic acid. Expression of the other aldehyde dehydrogenases was unaffected by the acetaldehyde addition.

CONCLUSIONS

The aldehyde dehydrogenases encoded by ALD2, ALD4 and ALD5 do not contribute to the elevated acetic acid production during Icewine fermentation. Expression of GPD2 was not upregulated in high sugar fermentations and does not reflect the elevated levels of glycerol found in these wines. Acetaldehyde at a concentration produced during Icewine fermentation stimulates the expression of ALD3, but has no impact on the expression of ALD2, -4, -5 and -6. Upregulation of ALD3 alone in the dilute fermentation is not sufficient to increase acetic acid in wine and requires additional responses found in cells under hyperosmotic stress.

SIGNIFICANCE AND IMPACT OF THE STUDY

This work confirms that increased acetic acid and glycerol production during Icewine fermentation follows upregulation of ALD3 and GPD1 respectively, but upregulation of ALD3 alone is not sufficient to increase acetic acid production. Additional responses of cells under osmotic stress are required to increase acetic acid in Icewine.

摘要

目的

我们之前报道过,基于发酵过程中这些基因的表达谱,由ALD3而非ALD6编码的醛脱氢酶部分导致了冰酒中乙酸含量的增加。我们现在已经完成了其余酵母醛脱氢酶基因ALD2、ALD4和ALD5在这些发酵过程中的表达谱分析,以确定它们对乙酸产生的贡献。研究了乙醛胁迫作为刺激这些发酵过程中醛脱氢酶(ALD)表达的信号对所有ALD基因的作用。在这些发酵过程中还跟踪了由GPD2编码的3-磷酸甘油的表达,以确定其作用,此外我们已经确定了GPD1在冰酒发酵过程中产生的甘油升高方面的作用。

方法与结果

使用商业葡萄酒酵母K1-V1116对冰酒汁(38.5°白利糖度,398±5 g l⁻¹糖)、稀释的冰酒汁(20.8°白利糖度,196±4 g l⁻¹糖)以及糖含量与原始冰酒汁相等的稀释汁(36.6°白利糖度,395±6 g l⁻¹糖)进行了重复发酵。与稀释汁发酵相比,冰酒发酵中乙酸和甘油的产量分别增加了8.4倍和2.7倍,在冰酒发酵条件下,第一周乙醛出现了四倍的短暂增加。由ALD4和ALD5编码的两种线粒体醛脱氢酶均有表达,在所有发酵开始时ALD5的表达最高,在每种条件的第一周ALD4的表达增加。ALD2、ALD4、ALD5和GPD2在三种发酵条件之间没有差异表达,表明它们与冰酒中乙酸和甘油的升高无关。当发酵稀释汁的酵母暴露于外源乙醛时,乙醛的短暂峰值增加了ALD3的表达,但仅此反应不足以导致乙酸增加。其他醛脱氢酶的表达不受乙醛添加的影响。

结论

由ALD2、ALD4和ALD5编码的醛脱氢酶对冰酒发酵过程中乙酸产量的升高没有贡献。GPD2的表达在高糖发酵中未上调,并且不能反映这些葡萄酒中甘油的升高水平。冰酒发酵过程中产生的浓度的乙醛刺激了ALD3的表达,但对ALD2、-4、-5和-6的表达没有影响。在稀释发酵中单独上调ALD3不足以增加葡萄酒中的乙酸,还需要细胞在高渗胁迫下的其他反应。

研究的意义和影响

这项工作证实了冰酒发酵过程中乙酸和甘油产量的增加分别是由于ALD3和GPD1的上调,但仅上调ALD3不足以增加乙酸产量。需要细胞在渗透胁迫下的其他反应来增加冰酒中的乙酸。

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