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利用微阵列技术对具有不同麦芽三糖利用谱的麦芽糖发酵酵母进行核型分析,揭示了参与麦芽糖和麦芽三糖利用的基因的拷贝数变化。

Microarray karyotyping of maltose-fermenting Saccharomyces yeasts with differing maltotriose utilization profiles reveals copy number variation in genes involved in maltose and maltotriose utilization.

机构信息

Departamento de Bioquímica, Universidade Federal de Santa Catarina, Florianópolis, SC, Brazil.

出版信息

J Appl Microbiol. 2010 Jul;109(1):248-59. doi: 10.1111/j.1365-2672.2009.04656.x. Epub 2009 Dec 18.

Abstract

AIMS

We performed an analysis of maltotriose utilization by 52 Saccharomyces yeast strains able to ferment maltose efficiently and correlated the observed phenotypes with differences in the copy number of genes possibly involved in maltotriose utilization by yeast cells.

METHODS AND RESULTS

The analysis of maltose and maltotriose utilization by laboratory and industrial strains of the species Saccharomyces cerevisiae and Saccharomyces pastorianus (a natural S. cerevisiae/Saccharomyces bayanus hybrid) was carried out using microscale liquid cultivation, as well as in aerobic batch cultures. All strains utilize maltose efficiently as a carbon source, but three different phenotypes were observed for maltotriose utilization: efficient growth, slow/delayed growth and no growth. Through microarray karyotyping and pulsed-field gel electrophoresis blots, we analysed the copy number and localization of several maltose-related genes in selected S. cerevisiae strains. While most strains lacked the MPH2 and MPH3 transporter genes, almost all strains analysed had the AGT1 gene and increased copy number of MALx1 permeases.

CONCLUSIONS

Our results showed that S. pastorianus yeast strains utilized maltotriose more efficiently than S. cerevisiae strains and highlighted the importance of the AGT1 gene for efficient maltotriose utilization by S. cerevisiae yeasts.

SIGNIFICANCE AND IMPACT OF THE STUDY

Our results revealed new maltotriose utilization phenotypes, contributing to a better understanding of the metabolism of this carbon source for improved fermentation by Saccharomyces yeasts.

摘要

目的

我们分析了 52 株能够高效发酵麦芽糖的酿酒酵母菌株对麦芽三糖的利用情况,并将观察到的表型与可能参与酵母细胞利用麦芽三糖的基因的拷贝数差异相关联。

方法和结果

我们使用微尺度液体培养以及好氧分批培养,对酿酒酵母和发酵毕赤酵母(一种天然的酿酒酵母/贝酵母杂种)的实验室和工业菌株的麦芽糖和麦芽三糖利用进行了分析。所有菌株都能有效地将麦芽糖作为碳源利用,但在利用麦芽三糖方面观察到三种不同的表型:高效生长、缓慢/延迟生长和不生长。通过微阵列核型分析和脉冲场凝胶电泳印迹,我们分析了选定酿酒酵母菌株中几种与麦芽糖相关基因的拷贝数和定位。虽然大多数菌株缺乏 MPH2 和 MPH3 转运蛋白基因,但几乎所有分析的菌株都具有 AGT1 基因和 MALx1 通透酶的拷贝数增加。

结论

我们的结果表明,发酵毕赤酵母菌株比酿酒酵母菌株更有效地利用麦芽三糖,并强调了 AGT1 基因对酿酒酵母高效利用麦芽三糖的重要性。

研究的意义和影响

我们的研究结果揭示了新的麦芽三糖利用表型,有助于更好地了解这种碳源的代谢,以提高酿酒酵母的发酵能力。

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