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Identification of target genes to control acetate yield during aerobic fermentation with Saccharomyces cerevisiae

机译:鉴定目标基因以控制酿酒酵母在好氧发酵过程中的乙酸盐产量

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Background Aerobic fermentation of grape must, leading to respiro-fermentative metabolism of sugars, has been proposed as way of reducing alcohol content in wines. Two factors limit the usefulness of Saccharomyces cerevisiae for this application, the Crabtree effect, and excess volatile acidity under aerobic conditions. This work aimed to explore the impact on ethanol acetate production of different S. cerevisiae strains deleted for genes previously related with the Crabtree phenotype. Results Recombinant strains were constructed on a wine industrial genetic background, FX10. All yeast strains, including FX10, showed respiro-fermentative metabolism in natural grape must under aerobic conditions, as well as a concomitant reduction in ethanol yield. This indicates that the Crabtree effect is not a major constrain for reaching relevant respiration levels in grape must. Indeed, only minor differences in ethanol yield were observed between the original and some of the recombinant strains. In contrast, some yeast strains showed a relevant reduction of acetic acid production. This was identified as a positive feature for the feasibility of alcohol level reduction by respiration. Reduced acetic acid production was confirmed by a thorough analysis of these and some additional deletion strains (involving genes HXK2 , PYK1 , REG1 , PDE2 and PDC1 ). Some recombinant yeasts showed altered production of glycerol and pyruvate derived metabolites. Conclusions REG1 and PDC1 deletion strains showed a strong reduction of acetic acid yield in aerobic fermentations. Since REG1 defective strains may be obtained by non-GMO approaches, these gene modifications show good promise to help reducing ethanol content in wines.
机译:背景技术已经提出,葡萄汁的需氧发酵会导致糖的呼吸发酵代谢,作为降低葡萄酒中酒精含量的方法。有两个因素限制了酿酒酵母在此应用中的用途,Crabtree效应和有氧条件下过量的挥发性酸度。这项工作旨在探讨删除以前与Crabtree表型有关的基因的啤酒酵母不同菌株对乙酸乙醇生产的影响。结果在葡萄酒工业遗传背景FX10上构建了重组菌株。所有酵母菌株(包括FX10)在有氧条件下必须在天然葡萄中表现出呼吸发酵代谢,同时乙醇产量也随之降低。这表明,Crabtree效应并不是达到葡萄汁中相关呼吸水平的主要限制。实际上,在原始菌株和一些重组菌株之间仅观察到乙醇产量的微小差异。相反,一些酵母菌株显示出乙酸产生的相关减少。这被认为是通过呼吸降低酒精水平的可行性的积极特征。通过对这些菌株和一些其他缺失菌株(涉及基因HXK2,PYK1,REG1,PDE2和PDC1)的彻底分析,确认了乙酸产量的降低。一些重组酵母显示甘油和丙酮酸衍生代谢产物的生产发生了变化。结论REG1和PDC1缺失菌株在好氧发酵中乙酸产量显着降低。由于可通过非转基因方法获得REG1缺陷菌株,因此这些基因修饰显示出有助于减少葡萄酒中乙醇含量的良好前景。

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