Tracking Trash to Treasure: In situ monitoring of Single Microbial Cell Oil biosynthesis from Waste Cooking Oil using Raman Reverse Isotope Probing and Imaging

Single microbial cell oil (SMCO) plays a fundamental role in maintaining the structure of cellular membranes and as energy reservoirs. Oleaginous microbes are known to have a high content of lipids, which often acts as a microbial factory for the bioremediation of waste cooking oil (WCO) which is a major pollutant contaminating land and water bodies. The biomass of microbes grown in WCO can be utilized to develop sustainable value-added products such as biodiesel, organic chemicals, bio preservatives, biosurfactants biopolymers. Conventional methods excel in SMCO analysis but lack efficacy for in situ or lysis-free monitoring of nascent SMCO synthesis and its turnover dynamics. To bridge this knowledge gap, in this study we have employed Raman reverse stable isotope probing (RrSIP) to investigate the time dependent nascent SMCO synthesis and assimilation in Yarrowia lipolytica, a well-known oleaginous yeast. Our finding provides a unique perspective for utilizing optical spectroscopy methods for lysis-free SMCO analysis which holds potential for utilization as an adjunct tool in bioprocess and biofuels industries.