Methanol-Dependent Carbon Fixation for Irreversible Synthesis of d-Allulose from d-Xylose by Engineered Escherichia coli.
Methanol-Dependent Carbon Fixation for Irreversible Synthesis of d-Allulose from d-Xylose by Engineered Escherichia coli.
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DOI：
10.1021/acs.jafc.2c06616
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发表时间：
2022-10
影响因子：
6.1
通讯作者：
Q. Guo;Mei-Ming Liu;Shang-He Zheng;Ling-Jie Zheng;Qian Ma;Ying-Kai Cheng;Su-ying Zhao;Li-Hai Fa
中科院分区：
文献类型：
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作者：
Q. Guo;Mei-Ming Liu;Shang-He Zheng;Ling-Jie Zheng;Qian Ma;Ying-Kai Cheng;Su-ying Zhao;Li-Hai Fa
d-Allulose因其甜度适中、能量低、具有独特的生理功能，是一种极具应用潜力的稀有己糖。目前的d-allulose生产策略，无论是工业化的还是正在开发的，利用六碳糖如d-葡萄糖或d-果糖作为底物，通常基于可逆的Izumoring外聚化原理。在这项工作中，我们设计了一种新的途径，将甲醇还原、戊糖磷酸（PP）、单磷酸核酮糖（RuMP）和单磷酸allulose （AuMP）的途径偶联在一起，使大肠杆菌从d-木糖和甲醇不可逆地合成d-allulose。通过SUMO融合提高AlsE表达，敲除FrmRAB、RpiA、PfkA、PfkB调节碳通量后，d-木糖补料发酵滴度达到≈70.7 mM， d-木糖的产率≈0.471 mM/mM，甲醇的产率≈0.512 mM/mM。
d-Allulose is a rare hexose with great application potential, owing to its moderate sweetness, low energy, and unique physiological functions. The current strategies for d-allulose production, whether industrialized or under development, utilize six-carbon sugars such as d-glucose or d-fructose as a substrate and are usually based on the principle of reversible Izumoring epimerization. In this work, we designed a novel route that coupled the pathways of methanol reduction, pentose phosphate (PP), ribulose monophosphate (RuMP), and allulose monophosphate (AuMP) for Escherichia coli to irreversibly synthesize d-allulose from d-xylose and methanol. After improving the expression of AlsE by SUMO fusion and regulating the carbon fluxes by knockout of FrmRAB, RpiA, PfkA, and PfkB, the titer of d-allulose in fed-batch fermentation reached ≈70.7 mM, with a yield of ≈0.471 mM/mM on d-xylose or ≈0.512 mM/mM on methanol.