Heterologous glucose-6-phosphate dehydrogenase empowers the biofuel potential of Tetradesmus obliquus via generating lipogenic NADPH

Abstract

Large-scale production of biofuels from oleaginous microalgal feedstocks in a commercially feasible manner is hampered by their relatively slow growth rates and overall lipid yield than the rapidly proliferating algal species. Therefore, it is pivotal to obviate this bottleneck by augmenting lipid overproduction in fast-growing microalgal species by heterologous overexpression lipogenic genes from model oleaginous species. In this regard, the pentose phosphate pathway of Tetradesmus obliquus was perturbed by heterologous overexpression of glucose-6-phosphate dehydrogenase (PtG6PD) from the model pennate diatom Phaeodactylum tricornutum, known for its oleaginicity. The role of PtG6PD in increasing lipid production was reported in the heterologous T. obliquus. Total lipid content was increased by 2.09- and 2.31-fold in transgenic lines than WT by elevating the lipogenic NADPH, without any negative impact on their growth rates. Interestingly, lipid yields in the transgenic strains reached up to 93 mg/L and 103 mg/L which showed a further 2-fold enhancement under nitrogen-deficient conditions. Furthermore, it was demonstrated that PtG6PD overexpression also altered fatty acid composition, with a specific increment in total monounsaturated fatty acids. Our results uncovered the pivotal role of the pentose phosphate pathway in supplying lipogenic NADPH in T. obliquus and identified key genetic targets for lipid biosynthesis enhancement in alternative hosts. Altogether, these findings open avenues for metabolic engineering in non-model and fast-growing non-oleaginous microalgal species, thereby presenting a promising strategy for empowering commercially viable photosynthetic cell factories for biofuel production.