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Molar-Based Targeted Metabolic Profiling of Cyanobacterial Strains with Potential for Biological Production

机译:基于摩尔的基于目标的蓝细菌菌株代谢分析及生物生产潜力

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摘要

Recently, cyanobacteria have become one of the most attractive hosts for biochemical production due to its high proliferative ability and ease of genetic manipulation. Several researches aimed at biological production using modified cyanobacteria have been reported previously. However, to improve the yield of bioproducts, a thorough understanding of the intercellular metabolism of cyanobacteria is necessary. Metabolic profiling techniques have proven to be powerful tools for monitoring cellular metabolism of various organisms and can be applied to elucidate the details of cyanobacterial metabolism. In this study, we constructed a metabolic profiling method for cyanobacteria using 13C-labeled cell extracts as internal standards. Using this method, absolute concentrations of 84 metabolites were successfully determined in three cyanobacterial strains which are commonly used as background strains for metabolic engineering. By comparing the differences in basic metabolic potentials of the three cyanobacterial strains, we found a well-correlated relationship between intracellular energy state and growth in cyanobacteria. By integrating our results with the previously reported biological production pathways in cyanobacteria, we found putative limiting step of carbon flux. The information obtained from this study will not only help gain insights in cyanobacterial physiology but also serve as a foundation for future metabolic engineering studies using cyanobacteria.
机译:近年来,由于蓝细菌的高增殖能力和易于遗传操作,它们已成为生化生产中最有吸引力的宿主之一。先前已经报道了一些针对使用修饰的蓝细菌进行生物生产的研究。但是,为了提高生物产品的产量,必须全面了解蓝细菌的细胞间代谢。代谢谱分析技术已被证明是监测各种生物体细胞代谢的强大工具,可用于阐明蓝细菌代谢的细节。在这项研究中,我们构建了以 13 C标记的细胞提取物为内标的蓝细菌代谢谱分析方法。使用这种方法,成功地确定了三种蓝细菌菌株中84种代谢物的绝对浓度,这些菌株通常用作代谢工程的背景菌株。通过比较三种蓝细菌菌株的基本代谢潜能的差异,我们发现细胞内能量状态与蓝细菌的生长之间存在良好相关的关系。通过将我们的结果与蓝细菌中先前报道的生物生产途径进行整合,我们发现了碳通量的推定限制步骤。从这项研究中获得的信息不仅将有助于对蓝细菌生理学的深入了解,而且还将为将来使用蓝细菌进行代谢工程研究奠定基础。

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