The development of synthetic microorganisms that could use one-carbon compounds, such as carbon dioxide, methanol, or formate, has received considerable interest. In this study, we engineered and to both synthetic methylotrophy and formatotrophy, enabling them to co-utilize methanol or formate with CO fixation through a synthetic C1-compound assimilation pathway (MFORG pathway). This pathway consisted of a methanol-formate oxidation module and the reductive glycine pathway. We first assembled the MFORG pathway in using endogenous enzymes, followed by blocking the native methanol assimilation pathway, modularly engineering genes of MFORG pathway, and compartmentalizing the methanol oxidation module. These modifications successfully enabled the methylotrophic yeast to utilize both methanol and formate. We then introduced the MFORG pathway from into the model yeast , establishing the synthetic methylotrophy and formatotrophy in this organism The resulting strain could also successfully utilize both methanol and formate with consumption rates of 20 mg/L/h and 36.5 mg/L/h, respectively. The ability of the engineered and to co-assimilate CO with methanol or formate through the MFORG pathway was also confirmed by C-tracer analysis. Finally, production of 5-aminolevulinic acid and lactic acid by co-assimilating methanol and CO was demonstrated in the engineered and . This work indicates the potential of the MFORG pathway in developing different hosts to use various one-carbon compounds for chemical production.

中文翻译：

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工程酵母共同利用甲醇或甲酸盐与二氧化碳固定


可以使用一碳化合物（例如二氧化碳、甲醇或甲酸盐）的合成微生物的开发引起了相当大的兴趣。在这项研究中，我们对合成甲基营养和格式营养进行了改造，使它们能够通过合成 C1 化合物同化途径（MFORG 途径）共同利用甲醇或甲酸与 CO 固定。该途径由甲醇-甲酸氧化模块和还原甘氨酸途径组成。我们首先使用内源酶组装MFORG途径，然后阻断天然甲醇同化途径，对MFORG途径基因进行模块化改造，并对甲醇氧化模块进行区隔化。这些修饰成功地使甲基营养酵母能够利用甲醇和甲酸。然后，我们将 MFORG 途径引入模型酵母中，在该生物体中建立合成甲基营养型和形态营养型。所得菌株还可以成功利用甲醇和甲酸，消耗率为 20 mg/L/h 和 36.5 mg/L/h，分别。 C-示踪剂分析也证实了工程化的和通过 MFORG 途径共同化 CO 与甲醇或甲酸盐的能力。最后，在工程化和 .这项工作表明了 MFORG 途径在开发不同宿主以使用各种一碳化合物进行化学生产方面的潜力。