FDH基因敲除和TsFDH转化提高大肠杆菌的生长速率
FDH knockout and TsFDH transformation lead to enhanced growth rate of Escherichia coli
摘要：
大气中二氧化碳含量超过400 ppm，导致全球气候异常。减少生物技术过程中释放的二氧化碳可以补救这些现象。在本研究中，我们试图找到一种解决方案，以减少在生长和繁殖过程中的二氧化碳量，通过防止甲酸转化为二氧化碳。甲酸到CO2的（生物）化学转化是关键反应。因此，我们比较了生长的BL 21，K12的一个亚家族，以及两个菌株，其中两个不同的基因相关的甲酸代谢被删除，在复杂和简单的媒体。实验结果与代谢预测完全一致。随后，敲除细菌比BL 21更有效地生长。有趣的是，TsFDH，一种具有将CO2转化为甲酸盐的倾向的甲酸盐脱氢酶，与没有TsFDH的细胞相比，增加了所有菌株的生长。大多数突变株在含有甘油的简单培养基中生长，这表明甘油是比葡萄糖更适合E.杆菌这些结果解释了以前的代谢模型预测不一致的原因，这些模型宣称甘油是大肠杆菌生长的合适碳源。但在实践中未能实现。为了对我们的观察进行更机理的评估，在E. coliRNA-seq数据集。基因表达相关结果显示，与蛋白质生物合成和甘油降解相关的几个基因的表达水平增加，这可能是我们观察到的结果的一个解释。
英文摘要：
Increased Atmospheric CO2 to over 400 ppm has prompted global climate irregularities. Reducing the released CO2 from biotechnological processes could remediate these phenomena. In this study, we sought to find a solution to reduce the amount of CO2 in the process of growth and reproduction by preventing the conversion of formic acid into CO2. The (bio)chemical conversion of formic acid to CO2 is a key reaction. Therefore, we compared the growth of BL21, being a subfamily of K12, alongside two strains in which two different genes related to the formate metabolism were deleted, in complex and simple media. Experimental results were entirely consistent with metabolic predictions. Subsequently, the knockout bacteria grew more efficiently than BL21. Interestingly, TsFDH, a formate dehydrogenase with the tendency of converting CO2 to formate, increased the growth of all strains compared with cells without the TsFDH. Most mutants grew in a simple medium containing glycerol, which showed that glycerol is the preferred carbon source compared to glucose for the growth of E. coli. These results explain the reasons for the inconsistency of predictions in previous metabolic models that declared glycerol as a suitable carbon source for the growth of E. coli but failed to achieve it in practice. To conduct a more mechanistic evaluation of our observations, RNA sequencing data analysis was conducted on an E. coli RNA-seq dataset. The gene expression correlation outcome revealed the increased expression levels of several genes related to protein biosynthesis and glycerol degradation as a possible explanation of our observations.
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