通过工程化大肠杆菌中的丙二酰辅酶 A 途径从葡萄糖中增强 3-羟基丙酸的生产。
Enhanced production of 3-hydroxypropionic acid from glucose via malonyl-CoA pathway by engineered Escherichia coli.
机构信息
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, China.
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, China; Shanghai Collaborative Innovation Center for Biomanufacturing Technology, 130 Meilong Road, Shanghai 200237, China.
出版信息
Bioresour Technol. 2016 Jan;200:897-904. doi: 10.1016/j.biortech.2015.10.107. Epub 2015 Nov 14.
In this study, production of 3-HP via malonyl-CoA was investigated by using metabolically engineered Escherichia coli carrying heterogeneous acetyl-CoA carboxylase (Acc) from Corynebacterium glutamicum and codon-optimized malonyl-CoA reductase (MCR) from Chloroflexus aurantiacus. Three engineered E. coli strains with different host-vector systems were constructed and investigated. The results indicated that the combination of E. coli BL21(DE3) and pET28a was the most efficient host-vector system for 3-HP production, and the highest concentration of 3-HP attained in shake flask cultivation reached 1.80g/L by the strain BE-MDA with induction at 0.25mM IPTG and 25°C, and supplementation of NaHCO3 and biotin. In fed-batch fermentation performed in a 5-L reactor, the concentration of 3-HP achieved 10.08g/L in 36h.
在这项研究中，通过使用携带来自谷氨酸棒杆菌的异源乙酰辅酶 A 羧化酶（Acc）和来自橙色硫细菌的密码子优化的丙二酰辅酶 A 还原酶（MCR）的代谢工程大肠杆菌来研究通过丙二酰辅酶 A 生产 3-HP。构建并研究了三种具有不同宿主载体系统的工程大肠杆菌菌株。结果表明，大肠杆菌 BL21(DE3)和 pET28a 的组合是用于 3-HP 生产的最有效的宿主载体系统，在摇瓶培养中，通过在 0.25mM IPTG 和 25°C以及补充 NaHCO3 和生物素的情况下诱导 BE-MDA 菌株，3-HP 的最高浓度达到 1.80g/L。在 5-L 反应器中进行的分批补料发酵中，36 小时内 3-HP 的浓度达到 10.08g/L。
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