Multi-Fold Enhancement of Tocopherol Yields Employing High CO₂ Supplementation and Nitrate Limitation in Native...

Tocopherols are the highly active form of the antioxidant molecules involved in scavenging of free radicals and protect the cell membranes from reactive oxygen species (ROS). In the present study, we focused on employing carbon supplementation with varying nitrate concentrations to enhance the total tocopherol yields in the native isolate Monoraphidium sp. CABeR41. The total tocopherol productivity of NR_HC (Nitrate replete + 3% CO₂) supplemented was (306.14 µg·L^-1 d^-1) which was nearly 2.5-fold higher compared to NR_VLC (Nitrate replete + 0.03% CO₂) (60.35 µg·L^-1 d^-1). The best tocopherol productivities were obtained in the NL_HC (Nitrate limited + 3% CO₂) supplemented cells (734.38 µg·L^-1 d^-1) accompanied by a significant increase in cell biomass (2.65-fold) and total lipids (6.25-fold). Further, global metabolomics using gas chromatography-mass spectrometry (GC-MS) was done in the defined conditions to elucidate the molecular mechanism during tocopherol accumulation. In the present study, the Monoraphidium sp. responded to nitrogen limitation by increase in nitrogen assimilation, with significant upregulation in gamma-Aminobutyric acid (GABA). Moreover, the tricarboxylic acid (TCA) cycle upregulation depicted increased availability of carbon skeletons and reducing power, which is leading to increased biomass yields along with the other biocommodities. In conclusion, our study depicts valorization of carbon dioxide as a cost-effective alternative for the enhancement of biomass along with tocopherols and other concomitant products like lipids and carotenoids in the indigenous strain Monoraphidium sp., as an industrial potential strain with relevance in nutraceuticals and pharmaceuticals.