Diatoms are eukaryotic microalgae that contain genes from various sources, including bacteria and the secondary endosymbiotic host. Due to this unique combination of genes, diatoms are taxonomically and functionally distinct from other algae and vascular plants and confer novel metabolic capabilities. Based on the genome annotation, we performed a genome-scale metabolic network reconstruction for the marine diatom Phaeodactylum tricornutum. Due to their endosymbiotic origin, diatoms possess a complex chloroplast structure which complicates the prediction of subcellular protein localization. Based on previous work we implemented a pipeline that exploits a series of bioinformatics tools to predict protein localization. The manually curated reconstructed metabolic network iLB1027_lipid accounts for 1,027 genes associated with 4,456 reactions and 2,172 metabolites distributed across six compartments. To constrain the genome-scale model, we determined the organism specific biomass composition in terms of lipids, carbohydrates, and proteins using Fourier transform infrared spectrometry. Our simulations indicate the presence of a yet unknown glutamine-ornithine shunt that could be used to transfer reducing equivalents generated by photosynthesis to the mitochondria. The model reflects the known biochemical composition of P. tricornutum in defined culture conditions and enables metabolic engineering strategies to improve the use of P. tricornutum for biotechnological applications.

Analysis of bulk biochemical composition is a key in fundamental and applied studies of microalgae and is essential to understanding responses to different cultivation scenarios. Traditional biochemical methods for the quantification of lipids, carbohydrates and proteins are often time-consuming, often involve hazardous reagents, require significant amounts of biomass and are highly dependent on practitioner proficiency. This study presents a rapid and non-destructive method, utilising Fourier-Transform Infrared (FTIR) spectroscopy for the simultaneous determination of lipid, protein and carbohydrate content in microalgal biomass. A simple univariate regression was applied to sets of reference microalgal spectra of known composition and recognised IR peak integrals. A robust single-species model was constructed, with coefficients of determination r(2)>0.95, high predictive accuracy and relative errors below 5%. The applicability of this methodology is demonstrated by monitoring the time-resolved changes in biochemical composition of the marine alga Nannochloropsis sp. grown to nitrogen starvation.

This work reports for the first time the detailed impacts of dual nitrogen (N) and phosphorus (P) stress on growth dynamics and biochemical composition in the Eustigmatophyte Nannochloropsis sp. P-stress concurrent with N-stress had subtle effects on culture bulk biochemical composition, but negatively influenced biomass productivity. However, the N:P supply ratio can be raised to at least 32:1 without compromising productivity (yielding a maximum lipid content of 52% of dry weight and volumetric lipid concentration of 233 mg L(-1)). The maximum biomass and lipid yields per unit of cell-P were 1.2 kg DW (gP)(-1) and 0.54 kg lipid (gP)(-1). The P concentration of many common media is thus in surplus for optimal Nannochloropsis sp. biomass and lipid production, offering potential for significant savings in P usage and improving the sustainability of algal cultivation.

This study monitored the biomass composition of

Microalgal biotechnology has yielded a range of products for different consumer markets, but large scale production for bulk commodities is limited by the cost and environmental impact of production. Nutrient requirements for large-scale production contribute significantly to the cost and environmental impact of microalgal biomass production and should subsequently be addressed by more careful sourcing of nutrients. This study assessed the use of nitrogen and phosphorus contained in effluents from anaerobic digestion of food waste to cultivate the marine microalga Nannochloropsis sp.. With suitable dilution, effluent could replace 100% of nitrogen demands and 16% of required phosphorus, without significant impacts on growth or biomass productivity. Additional phosphorus requirements could be decreased by increasing the N:P molar ratio of the media from 16:1 to 32:1. Nannochloropsis sp. accumulated lipid up to 50% of dry weight under N-stress, with significant increases in the content of saturated and mono-unsaturated fatty acids. Using empirical data generated in this study, the cost and environmental impact of nitrogen and phosphorus supply was assessed versus the use of fertilizers for biomass and biodiesel production. Nutrient requirements predicted by the Redfield Ratio overestimating impacts by as much as 140% compared to empirical data. By utilising residual nutrients and optimising nutrient supply, the cost and environmental impact of nitrogen and phosphorus were decreased by 90% versus the use of artificial fertilizers.