Attenuated total reflection–Fourier transform infrared (ATR–FTIR) spectroscopy is a simple, cheap, and fast method to collect chemical compositional information from microalgae. However, (semi)quantitative evaluation of the collected data can be daunting. In this work, ATR–FTIR spectroscopy was used to monitor changes of protein, lipid, and carbohydrate content in seven green microalgae grown under nitrogen starvation. Three statistical methods—univariate linear regression analysis (ULRA), orthogonal partial least squares (OPLS), and multivariate curve resolution-alternating least squares (MCR–ALS)—were compared in their ability to model and predict the concentration of these compounds in the biomass. OPLS was found superior, since it i) included all three compounds simultaneously; ii) explained variations in the data very well; iii) had excellent prediction accuracy for proteins and lipids, and acceptable for carbohydrates; and iv) was able to discriminate samples based on cultivation stage and type of storage compounds accumulated in the cells. ULRA models worked well for the determination of proteins and lipids, but carbohydrates could only be estimated if already determined protein contents were used for scaling. Results obtained by MCR–ALS were similar to ULRA, however, this method is considerably easier to perform and interpret than the more abstract statistical/chemometric methods. FTIR-spectroscopy-based models allow high-throughput, cost-effective, and rapid estimation of biomass composition of green microalgae.

Rapid rises in atmospheric CO₂ levels derived from fossil fuel combustion are imposing urgent needs for renewable substitutes. One environmentally friendly alternative is biodiesel produced from suitable microalgal fatty acids. Algal strains normally grow photoautotrophically, but this is problematic in Northern areas because of the light limitations for much of the year. Mixotrophic and particularly heterotrophic strains could be valuable, especially if they can be cultivated in municipal wastewater with contents of nutrients such as nitrogen and phosphorous that should be reduced before release into receiving water. Thus, the aim of this study was to screen for microalgal strains suitable for heterotrophic cultivation with a cheap carbon source (glycerol) for biodiesel production in Nordic, and other high-latitude, countries. One of the examined strains, a Desmodesmus sp. strain designated 2-6, accumulated biomass at similar rates in heterotrophic conditions with 40 mM glycerol as in autotrophic conditions. Furthermore, in heterotrophic conditions it produced more fatty acids, and ca. 50% more C18:1 fatty acids, as well as showing a significant decrease in C18:3 fatty acids, all of which are highly desirable features for biodiesel production.

In order to investigate environmentally sustainable sources of organic carbon and nutrients, four Nordic green microalgal strains, Chlorella sorokiniana, Chlorella saccharophila, Chlorella vulgaris, and Coelastrella sp., were grown on a wood (Silver birch, Betula pendula) hydrolysate and dairy effluent mixture. The biomass and lipid production were analysed under mixotrophic, as well as two-stage mixotrophic/heterotrophic regimes. Of all of the species, Coelastrella sp. produced the most total lipids per dry weight (~40%) in the mixture of birch hydrolysate and dairy effluent without requiring nutrient (nitrogen, phosphorus, and potassium—NPK) supplementation. Overall, in the absence of NPK, the two-stage mixotrophic/heterotrophic cultivation enhanced the lipid concentration, but reduced the amount of biomass. Culturing microalgae in integrated waste streams under mixotrophic growth regimes is a promising approach for sustainable biofuel production, especially in regions with large seasonal variation in daylight, like northern Sweden. To the best of our knowledge, this is the first report of using a mixture of wood hydrolysate and dairy effluent for the growth and lipid production of microalgae in the literature.