Non-natural Cofactor and Formate-Driven Reductive Carboxylation of Pyruvate
Non-natural Cofactor and Formate-Driven Reductive Carboxylation of Pyruvate
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非天然辅因子和甲酸盐驱动的丙酮酸还原羧化
DOI：
10.1002/anie.201915303
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发表时间：
2020-01-21
影响因子：
16.6
通讯作者：
Zhao, Zongbao K.
中科院分区：
文献类型：
--
作者：
Guo, Xiaojia;Liu, Yuxue;Zhao, Zongbao K.
提出了一种用于丙酮酸还原羧化的非天然辅因子和甲酸盐驱动的系统。获得了甲酸脱氢酶 (FDH) 突变体 FDH*，它有利于非天然氧化还原辅因子烟酰胺胞嘧啶二核苷酸 (NCD)，以牺牲甲酸为代价生成专用还原当量。通过耦合 FDH* 和 NCD 依赖性苹果酸酶 (ME*)，证明了甲酸的成功利用作为丙酮酸还原羧化的 CO2 源和电子供体，并且甲酸和苹果酸之间具有完美的化学计量。当 C-13 同位素标记的甲酸用于体外试验时，高达 53% 的苹果酸具有标记的碳原子。在模型宿主大肠杆菌中表达 FDH* 和 ME* 后，工程菌株在存在甲酸和 NCD 的情况下产生更多的苹果酸。这项工作为二氧化碳固定提供了一种替代的原子经济策略，其中使用甲酸盐代替二氧化碳并提供专用的还原能力。
A non-natural cofactor and formate driven system for reductive carboxylation of pyruvate is presented. A formate dehydrogenase (FDH) mutant, FDH*, that favors a non-natural redox cofactor, nicotinamide cytosine dinucleotide (NCD), for generation of a dedicated reducing equivalent at the expense of formate were acquired. By coupling FDH* and NCD-dependent malic enzyme (ME*), the successful utilization of formate is demonstrated as both CO2 source and electron donor for reductive carboxylation of pyruvate with a perfect stoichiometry between formate and malate. When C-13-isotope-labeled formate was used in in vitro trials, up to 53 % of malate had labeled carbon atom. Upon expression of FDH* and ME* in the model host E. coli, the engineered strain produced more malate in the presence of formate and NCD. This work provides an alternative and atom-economic strategy for CO2 fixation where formate is used in lieu of CO2 and offers dedicated reducing power.