Molar-based targeted metabolic profiling of cyanobacterial strains with potential for biological production.
Molar-based targeted metabolic profiling of cyanobacterial strains with potential for biological production.
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DOI：
10.3390/metabo4020499
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发表时间：
2014-06-20
期刊：
影响因子：
4.1
通讯作者：
Fukusaki E
中科院分区：
文献类型：
--
作者：
Dempo Y;Ohta E;Nakayama Y;Bamba T;Fukusaki E
近年来，蓝藻因其增殖能力强、易于遗传操作等优点，已成为最具吸引力的生化生产宿主之一。以前已经报道了几项旨在使用修饰的蓝藻进行生物生产的研究。然而，为了提高生物产品的产量，深入了解蓝藻的细胞间代谢是必要的。代谢分析技术已被证明是监测各种生物体细胞代谢的有力工具，并可用于阐明蓝藻代谢的细节。在这项研究中，我们构建了一个代谢分析方法，蓝藻使用13 C-标记的细胞提取物作为内标。利用该方法成功地测定了3株代谢工程背景菌株中84种代谢产物的绝对浓度。通过比较三种蓝藻基本代谢势的差异，发现蓝藻细胞内能量状态与生长之间存在良好的相关性。通过将我们的结果与先前报道的蓝藻生物生产途径相结合，我们发现了假定的碳通量限制步骤。从这项研究中获得的信息不仅有助于获得蓝藻生理学的见解，而且还为未来利用蓝藻进行代谢工程研究奠定了基础。
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.