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应用拉曼光谱学结合奇异值分解方法在单细胞尺度上对杀虫贪铜菌(C.necator)H16菌株在不同的氮源下合成聚β-羟基丁酸(PHB)的代谢动态进行分析。结果显示,硫酸铵是PHB发酵的理想氮源,对应PHB发酵速度、效率和产量在测试氮源中表现最好。奇异值分解结果显示,源自RNA、DNA、蛋白质和PHB的拉曼峰是发酵的主要特征,随着发酵进程的延伸,菌体细胞差异、产物含量差别逐渐增大。分析PHB产物快速合成过程,可见表征核酸、蛋白质和PHB的782、1574、1660、1732 cm~(-1)等峰的强度变化活跃;不同氮源下,782 cm~(-1)峰与1660 cm~(-1)峰的强度呈正相关,而1660 cm~(-1)峰与1732 cm~(-1)峰的强度呈负相关。因此,不同氮源可能影响细胞的RNA代谢和蛋白质代谢,从而间接影响PHB的合成。拉曼光谱结合数据挖掘技术可以分析微生物发酵过程中的代谢信息,从分子光谱的角度为寻找最佳的发酵条件提供新的信息。
Raman spectroscopy combined with singular value decomposition was used to analyze the metabolic kinetics of poly-β-hydroxybutyrate (PHB) synthesized by C. necator strain H16 under different nitrogen sources on a single-cell scale. The results showed that ammonium sulfate is the ideal nitrogen source for PHB fermentation. Corresponding to the PHB fermentation rate, the efficiency and yield are the best among the tested nitrogen sources. The results of singular value decomposition showed that the Raman peaks originated from RNA, DNA, protein and PHB are the main characteristics of fermentation. With the extension of fermentation process, the difference of cells and the increase of product content gradually increase. The rapid synthesis of PHB products was analyzed. The results showed that the peaks of 782, 1574, 1660 and 1732 cm ~ (-1) which characterize nucleic acids, proteins and PHB were active. The peak of 782 cm ~ (-1) The cm ~ (-1) peak intensity was positively correlated, while the peak intensity of 1660 cm ~ (-1) peak was negatively correlated with the intensity of 1732 cm ~ (-1) peak. Therefore, different nitrogen sources may affect the cell’s RNA metabolism and protein metabolism, which indirectly affect the PHB synthesis. Raman spectroscopy combined with data mining techniques can be used to analyze metabolic information during microbial fermentation and provide new information for finding the best fermentation conditions from molecular spectroscopy.