The high demand for sustainable biodiesel feedstocks has led to the exploration of innovative strategies to enhance lipid productivity in microalgae. This study introduces a push–pull strategy to optimize lipid accumulation in Nannochloropsis oculata. The benzyl amino purine (BAP) and naphthalene acetic acid (NAA) stimulation, acting as the ‘push’ component, significantly boost growth and nutrient stress tolerance. Meanwhile, the ‘pull’ component, nitrogen (N) deficiency, triggers lipid biosynthesis. A Box–Behnken design was employed to optimize the factors named BAP fraction (0–1), total phytohormone (PH) BAP/NAA mix dose (0–20 ppm), and N-concentration (0–50%). The combined BAP/NAA treatment significantly increased biomass (15% higher than the control) and mitigated N-stress with higher doses (20 ppm). Lipid yield surged from 12.4% to 38.87% under optimized conditions (23.25% N, 39.5 ppm NAA, and BAP fraction 0). The push–pull strategy contributed to boosting lipid synthesis and balancing biomass production. N-limitation and total PH dosage were the determining factors in this strategy. This work demonstrates the potential of the push–pull strategy in increasing lipid accumulation, offering a promising and optimistic solution for biodiesel production at scale from microalgae. By reducing dependence on fossil fuels, N. oculata emerges as a reliable feedstock for oil extraction and biodiesel.

Plant enzymes, such as those from castor bean (Ricinus communis L.), have been studied as biocatalysts for various industrial applications. In this study, a powdered enzyme extract was used to analyze the potential of lipases present in three castor bean varieties (LV1–LV3). Their hydrolytic activity was evaluated against olive and castor oil substrates (refined and non-refined) at pH 4.5 and 7.0. The results showed that LV2 had higher catalytic activity, managing to hydrolyze more than 98% of the oils in only 4.5 h of reaction. Additionally, diverse computational tools were employed to assess the behavior of OBL1 lipase. By molecular docking analysis, it was found that the binding of ricinoleic acid triacylglycerols to the catalytic site was favored at an acidic pH of 4.5 due to enhanced hydrogen bond formation. In contrast, at pH 7.0, hydrophobic interactions predominated. Molecular dynamics analyses of OBL1 showed that the enzyme–substrate complex is stable over time, providing a robust molecular explanation for the high catalytic efficiency observed in the experimental assays. This research stands out as one of the first to combine lab experiments with computational modeling to study castor bean lipases. Our integrated approach provides fresh insights into the enzyme’s versatile catalytic capabilities, underscoring its potential as a powerful and reliable tool for industrial biotechnology.

The high demand for sustainable biodiesel feedstocks has led to the exploration of innovative strategies to enhance lipid productivity in microalgae. This study introduces a push-pull strategy to optimize lipid accumulation in Nannochloropsis oculata. The benzyl amino purine (BAP) and naphthalene acetic acid (NAA) stimulation, acting as the 'push' component, significantly boost growth and nutrient-stress tolerance. Meanwhile, the 'pull' component, nitrogen (N) deficiency, triggers lipid biosynthesis. A Box-Behnken design was employed to optimize the factors named BAP fraction (0-1), total phytohormone (PH) BAP/NAA mix dose (0-20 ppm), and N-concentration (0-50%). The combined BAP/NAA treatment not only significantly increased biomass (15% higher than the control) but also mitigated N-stress with higher doses (20 ppm). Lipid yield surged from 12.4% to 38.87% under optimized conditions (23.25% N, 39.5 ppm NAA, and BAP fraction 0). The push-pull strategy contributed to boosting lipid synthesis and balancing biomass production. N-limitation and total PH dosage were the determining factors in this strategy. This work demonstrates the potential of the push-pull strategy in increasing lipid accumulation, offering a promising and optimistic solution for biodiesel production at scale from microalgae. By reducing dependence on fossil fuels, N. oculata emerges as a confident feedstock for oil extraction and biodiesel.