Global Metabolic Engineering of Glycolytic Pathway via Multicopy Integration in Saccharomyces cerevisiae.
Global Metabolic Engineering of Glycolytic Pathway via Multicopy Integration in Saccharomyces cerevisiae.
复制标题
通过酿酒酵母多拷贝整合进行糖酵解途径的全局代谢工程。
DOI：
10.1021/acssynbio.6b00281
复制
发表时间：
2017
影响因子：
4.7
通讯作者：
H. Ogino
中科院分区：
文献类型：
--
作者：
Ryosuke Yamada;Kazuki Wakita;H. Ogino
通过改造酿酒酵母的代谢途径，使用可再生原料生产生物燃料和生物基化学品最近已成为一种有吸引力的选择。许多研究人员试图通过过度表达一些糖酵解酶来提高葡萄糖消耗率，因为大多数目标生物基化学品都是通过糖酵解衍生的。然而，这些尝试收效甚微。在这项研究中，为了创建具有高葡萄糖消耗率的酿酒酵母菌株，我们使用多拷贝整合来开发全局代谢工程策略。在大约 350 个代谢工程菌株中，YPH499/dPdA3-34 表现出最高的葡萄糖消耗率。该菌株的细胞生长速度和葡萄糖消耗速度比对照菌株高1.3倍。实时PCR分析显示，YPH499/dPdA3-34中糖酵解相关基因（如HXK2、PFK1、PFK2、PYK2、PGI1和PGK1）的转录水平增加。因此，我们的策略是优化酿酒酵母全局代谢途径的一种有前途的方法。
The use of renewable feedstocks for producing biofuels and biobased chemicals by engineering metabolic pathways of yeast Saccharomyces cerevisiae has recently become an attractive option. Many researchers attempted to increase glucose consumption rate by overexpressing some glycolytic enzymes because most target biobased chemicals are derived through glycolysis. However, these attempts have met with little success. In this study, to create a S. cerevisiae strain with high glucose consumption rate, we used multicopy integration to develop a global metabolic engineering strategy. Among approximately 350 metabolically engineered strains, YPH499/dPdA3-34 exhibited the highest glucose consumption rate. This strain showed 1.3-fold higher cell growth rate and glucose consumption rate than the control strain. Real-time PCR analysis revealed that transcription levels of glycolysis-related genes such as HXK2, PFK1, PFK2, PYK2, PGI1, and PGK1 in YPH499/dPdA3-34 were increased. Our strategy is thus a promising approach to optimize global metabolic pathways in S. cerevisiae.