Revealing Lipid Body Formation and its Subcellular Reorganization in Oleaginous Microalgae Using Correlative Optical Microscopy and Infrared Nanospectroscopy Atomic Force Microscopy-Induced Resonance (AFM-IR)

The purpose of this work is to develop an integrated imaging approach to characterize without labeling at the sub-cellular level the formation of lipid body droplets (LBs) in microalgae undergoing nitrogen starvation. First conventional optical microscopy approaches, gas chromatography, and turbidimetry measurements allowed to monitor the biomass and the total lipid content in the oleaginous microalgae Parachlorella kesslerii during the starvation process. Then a local analysis of the LBs was proposed using an innovative infrared nanospectroscopy technique called atomic force microscopy-based infrared spectroscopy (AFM-IR). This label-free technique assessed the formation of LBs and allowed to look into the LB composition thanks to the acquisition of local infrared spectra. Last correlative measurements using fluorescence microscopy and AFM-IR were performed to investigate the subcellular reorganization of LB and the chloroplasts.

Biodiesel Production From Lignocellulosic Biomass Using Oleaginous Microbes: Prospects for Integrated Biofuel Production

Biodiesel is an eco-friendly, renewable, and potential liquid biofuel mitigating greenhouse gas emissions. Biodiesel has been produced initially from vegetable oils, non-edible oils, and waste oils. However, these feedstocks have several disadvantages such as requirement of land and labor and remain expensive. Similarly, in reference to waste oils, the feedstock content is succinct in supply and unable to meet the demand. Recent studies demonstrated utilization of lignocellulosic substrates for biodiesel production using oleaginous microorganisms. These microbes accumulate higher lipid content under stress conditions, whose lipid composition is similar to vegetable oils. In this paper, feedstocks used for biodiesel production such as vegetable oils, non-edible oils, oleaginous microalgae, fungi, yeast, and bacteria have been illustrated. Thereafter, steps enumerated in biodiesel production from lignocellulosic substrates through pretreatment, saccharification and oleaginous microbe-mediated fermentation, lipid extraction, transesterification, and purification of biodiesel are discussed. Besides, the importance of metabolic engineering in ensuring biofuels and biorefinery and a brief note on integration of liquid biofuels have been included that have significant importance in terms of circular economy aspects.

Nutrient deficiency and an algicidal bacterium improved the lipid profiles of a novel promising oleaginous dinoflagellate, Prorocentrum donghaiense, for biodiesel production

The lipid production potential of 8 microalgae species was investigated. Among these eight species, the best strain was a dominant bloom-causing dinoflagellate, Prorocentrum donghaiense ; this species had a lipid content of 49.32±1.99% and exhibited a lipid productivity of 95.47±0.99 mg L −1 d −1 , which was 2-fold higher than the corresponding values obtained for the oleaginous microalgae Nannochloropsis gaditana and Phaeodactylum tricornutum . P. donghaiense, which is enriched in C16:0 and C22:6, is appropriate for commercial DHA production. Nitrogen or phosphorus stress markedly induced lipid accumulation to levels surpassing 75% of the dry weight, increased the C18:0 and C17:1 contents, and decreased the C18:5 and C22:6 contents, and these effects resulted in decreases in the unsaturated fatty-acid levels and changes in the lipid properties of P. donghaiense such that the species met the biodiesel specification standards. Compared with the results obtained under N-deficient conditions, the enhancement in the activity of alkaline phosphatase of P. donghaiense observed under P-deficient conditions could partly alleviate the adverse effects on the photosynthetic system exerted by P deficiency to induce the production of more carbohydrates for lipogenesis. The supernatant of the algicidal bacterium Paracoccus sp. Y42 culture lysed P. donghaiense without decreasing its lipid content, which resulted in facilitation of the downstream oil extraction process and energy savings through the lysis of algal cells. The Y42 supernatant treatment improved the lipid profiles of algal cells by increasing their C16:0, C18:0 and C18:1 contents and decreasing their C18:5 and C22:6 contents, which is favourable for biodiesel production. IMPORTANCE This study demonstrates the high potential of P. donghaiense , a dominant bloom-causing dinoflagellate, for lipid production. Compared with previously studied oleaginous microalgae, P. donghaiense exhibit greater potential for practical application due to its higher biomass and lipid contents. Nutrient deficiency and the algicidal bacterium Paracoccus sp. Y42 could improve the suitability of the lipid profile of P. donghaiense for biodiesel production. Furthermore, Paracoccus sp. Y42 effectively lyse algal cells, which facilitates the downstream oil extraction process for biodiesel production and results in energy savings through the lysing of algal cells. This study provides a more promising candidate for the production of DHA for human nutritional products and of microalgal biofuel, as well as a more cost-effective method for breaking algal cells. The high lipid productivity of P. donghaiense and algal cell lysis by algicidal bacteria contribute to reductions in the production cost of microalgal oil.

Effects of Laser Mutagenesis on Microalgae Production and Lipid Accumulation in Two Economically Important Fresh Chlorella Strains under Heterotrophic Conditions

Microalgae produce a variety of high-value products. Enhancing product contents in microalgal cells is one of the efficient ways to decrease production costs. Improved germplasm and heterotrophic cultivation may enhance microalgae biomass and lipid content. In this study, we investigated the effect of three types of laser irradiation and heterotrophic cultivation on lipid productivity, lipid content, and biomass of two Chlorella strains (i.e., FACHB 9 and FACHB 31). Results showed that the highest biomasses of 4.81 g/L (15.03-fold) and 4.66 g/L (7.32-fold) were obtained in the third generation of FACHB 9 and FACHB 31 induced by a neodymium-doped yttrium aluminum garnet (Nd:YAG) laser for 8 min and 12 min, respectively. The highest lipid contents were 525.6 mg/g (1.67-fold) dry weight (DW) and 780.0 mg/g DW (2.20-fold) in the third and the first generations of FACHB 9 and FACHB 31 induced by Nd:YAG for 8 min and by a helium–neon (He–Ne) laser for 4 min, respectively. The highest lipid productivities of 69.82 ± 3.29 mg/L/d (19.7-fold) and 30.71 ± 3.77 mg/L/d (3.1-fold) were obtained in FACHB 9 and FACHB 31 treated by a semiconductor (SC) laser for 4 min and by a He–Ne laser for 12 min, respectively. Our study suggested that laser mutagenesis is a potential method for screening economically important oleaginous microalgae strains.