Methanol is increasingly becoming an attractive carbon feedstock for the production of various biochemicals due to its high abundance and low price. In this study, when methanol assimilation module was introduced into succinic acid producing Escherichia coli by employing the NAD-dependent methanol dehydrogenase from Bacillus methanolicus and ribulose monophosphate pathway from different donor organisms, succinic acid yield was increased from 0.91 ± 0.08 g/g to 0.98 ± 0.11 g/g with methanol as an auxiliary substrate under the anaerobic fermentation. Further ¹³C-labeling experiments showed that the recombinant strain successfully converted methanol into succinic acid, as the carbon atom of carboxyl group in succinic acid was labeled by ¹³C. It was found that the NADH generated by methanol oxidation would benefit succinate production, as the NADH/NAD⁺ ratio in vivo was decreased from 0.67 to 0.45 in the engineered strain, indicating that the efficiency of succinic acid synthesis was significantly improved when driven by methanol. This study represents a successful case for the development of reducing chemical production using methanol as an auxiliary substrate.

中文翻译：

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甲醇代谢琥珀酸高产生产的大肠杆菌代谢工程

甲醇由于其丰度高和价格低廉，正日益成为用于生产各种生物化学品的有吸引力的碳原料。在这项研究中，当通过使用甲醇芽孢杆菌的NAD依赖性甲醇脱氢酶和来自不同供体生物体的核糖单磷酸途径将甲醇同化模块引入产琥珀酸的大肠杆菌中时，琥珀酸的产率从0.91±0.08 g / g增加到0.98在厌氧发酵条件下，以±0.11 g / g的甲醇为辅助底物。进一步的¹³ C标记实验表明，该重组菌株成功地将甲醇转化为琥珀酸，因为琥珀酸中羧基的碳原子被¹³标记。C.发现通过甲醇氧化产生的NADH将有利于琥珀酸的生产，因为在工程菌株中体内的NADH / NAD ⁺比率从0.67降低到0.45，表明驱动时琥珀酸的合成效率得到了显着提高。用甲醇。这项研究代表了使用甲醇作为辅助底物开发减少化学生产的成功案例。