Karlo Jiro, Dhillon Ashish Kumar, Siddhanta Soumik, Singh Surya Pratap
Department of Biosciences and Bioengineering, Indian Institute of Technology Dharwad, Dharwad, India.
Department of Chemistry, Indian Institute of Technology Delhi, New Delhi, India.
J Biophotonics. 2024 Feb;17(2):e202300341. doi: 10.1002/jbio.202300341. Epub 2023 Dec 4.
Global proteome changes in microbes affect the survival and overall production of commercially relevant metabolites through different bioprocesses. The existing methods to monitor proteome level changes are destructive in nature. Stable isotope probing (SIP) coupled with Raman spectroscopy is a relatively new approach for proteome analysis. However, applying this approach for monitoring changes in a large culture volume is not cost-effective. In this study, for the first time we are presenting a novel method of combining reverse SIP using C-glucose and Deuterium to monitor the proteome changes through Raman spectroscopy. The findings of the study revealed visible changes (blue shifts) in proteome related peaks that can be used for monitoring proteome dynamics, that is, synthesis of nascent amino acids and its turnover with time in a non-destructive, cost-effective, and label-free manner.
微生物中的全局蛋白质组变化通过不同的生物过程影响商业相关代谢物的存活和整体产量。现有的监测蛋白质组水平变化的方法本质上具有破坏性。稳定同位素探测（SIP）与拉曼光谱联用是一种相对较新的蛋白质组分析方法。然而，将这种方法应用于监测大量培养物中的变化并不具有成本效益。在本研究中，我们首次提出了一种结合使用¹³C-葡萄糖和氘的反向SIP通过拉曼光谱监测蛋白质组变化的新方法。该研究结果揭示了蛋白质组相关峰中可见的变化（蓝移），可用于以非破坏性、具有成本效益且无标记的方式监测蛋白质组动态，即新生氨基酸的合成及其随时间的周转。
