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使用mBFP在大肠杆菌中酸通路中识别了一种新的NADPH生成反应

Koichiro Ueno¹, Shogo Sawada¹, Mai Ishibashi¹

¹Department of Bioinformatic Engineering, Graduate School of Information Science and Technology, Osaka University, Suita, Osaka, Japan.

Journal of bacteriology

|October 10, 2024

概括

研究人员发现了一种新的方法,使大肠杆菌使用西洛斯产生NADPH,确定葡萄糖-6-酸盐脱酶作为关键酶. 这一发现提供了一种代谢工程策略,用于从西洛中产生有价值的化合物.

科学领域:

微生物学微生物学
生物化学生物化学
代谢工程是代谢工程.

背景情况:

NADPH对于合成代谢反应至关重要,但其在大肠杆菌中的生成途径尚未完全理解.
识别新的NADPH生成路径对于代谢工程应用至关重要.

研究的目的:

探索和识别E. coli中以前未知的NADPH生成反应.
为了研究NADPH生产,使用树脂素作为碳源.

主要方法:

利用基因组衍生的蓝色光蛋白 (mBFP) 作为细胞内NADPH记者来监测光动态.
对糖-6-酸盐异构酶 (PGI) 删除菌株及其父菌株进行了扰动分析.
用纯化的酶进行了体外检测,以确认基质利用.

主要成果:

观察到与西洛斯的ΔPGI菌株中NADPH生成的增加,这表明了一个意想不到的途径.
鉴定了葡萄糖-6-酸脱酶 (G6PDH) 和6-酸酸脱酶 (6PGDH) 作为在化物条件下对NADPH生产的贡献者.
证明G6PDH可以利用红-4-酸盐 (E4P) 产生NADPH,尽管通常在细胞内度较低.

结论:

在大肠杆菌中发现了一种新的NADPH生成途径,其中涉及g6pdh和e4p在西洛斯代谢过程中.
这条通路的流量随着细胞内E4P升高而增加,提供了代谢工程目标.
提供了一种增强NADPH再生的策略,使用西洛斯生物生产有价值的化合物.

关键词:

埃舍里希亚大肠杆菌 (Escherichia coli) 是一个大肠杆菌.在 G6PDH 中.它们是NADPH,NADPH.在 mBFP mBFP.饥饿饥饿饥饿饥饿饥饿一个Xylose,一个Xylose.

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