Commercial production of microalgal biodiesel is not yet economically viable, largely because of low storage lipid yield in microalgae mass cultivation. Selection of lipid-rich microalgae, thus, becomes one of the key research topics for microalgal biodiesel production. However, the laboratory screening protocols alone cannot predict the ability of the strains to dominate and perform in outdoor ponds. Comprehensive assessment of microalgae species should be performed not only under the laboratory conditions, but also in the fields. Laboratory investigations using a bubbled column photobioreactor indicated the microalga Graesiella sp. WBG-1 to be the most productive species among the 63 Chlorophyta strains. In a 10 L reactor, mimicking the industrial circular pond, Graesiella sp. WBG-1 produced 12.03 g biomass m−2 day−1 and 5.44 g lipids (45.23 % DW) m−2 day−1 under 15 mol m−2 day−1 artificial light irradiations. The lipid content decreased to ~34 % DW when the microalga was cultured in 30 L tank PBR under natural solar irradiations, but the decline of lipid content with scaling up was the minimum among the tested strains. Based on these results, the microalga was further tested for its lipid production and culture competitiveness using a pilot-scale raceway pond (200 m2 illuminated area, culture volume 40,000 L). Consequently, Graesiella sp. WBG-1 maintained a high lipid content (33.4 % DW), of which ~90 % was storage TAGs. Results from the outdoor experiments indicated the nice adaptability of the Graesiella sp. WBG-1 to strong and fluctuating natural solar irradiance and temperature, and also demonstrated several other features, such as large cell size (easy for harvest and resistant to swallow by protozoa) and tolerance to high culture pH (helpful to CO2 fixation). Graesiella sp. WBG-1 was a promising strain capable of accumulating large amount of storage lipid under nature solar irradiance and temperature. The high lipid content of 33.4 % DW was achieved for the first time in pilot-scale raceway pond. The results also provide evidence for the feasibility of using low-cost raceway pond for autotrophic cultivation of microalgae for biodiesel production.

The water-soluble pigment protein phycocyanin (C-PC) from cyanobacteria Arthrospira sp. is an excellent natural food colorant and nutritional supplement with a brilliant blue color. However, C-PC is highly unstable, especially at high temperatures and when exposed to oxidative stress. The lack of simple and economical methods for improving the stability of C-PC greatly limits the application of this functional protein in the food industry. This study investigated the effect of adding saccharides (glucose, mannose, galactose, and maltose) and sugar alcohols (mannitol and maltitol) on the stability of food-grade C-PC extracted from Arthrospira platensis; the relevant reaction kinetics were also analyzed. The results revealed that glucose, mannose, mannitol, galactose, and maltose could effectively improve the thermal stability of C-PC. This improvement was positively correlated with the concentration of the additives and decreased sharply when the temperature exceeded 60 °C. Furthermore, the results also revealed the instability of C-PC when subjected to oxidative stress and the effectiveness of glucose, mannose, mannitol, and maltose in preventing the oxidative degradation of C-PC. In general, this study demonstrates that glucose, mannose, mannitol, and maltose are promising compounds for promoting the thermal and oxidative stability of C-PC, providing an economical and effective method for C-PC preservation.

Microalgae are an important feedstock in industries. Currently, efforts are being made in the non-phototrophic cultivation of microalgae for biomass production. Studies have shown that mixotrophy is a more efficient process for producing algal biomass in comparison to phototrophic and heterotrophic cultures. However, cultivation of microalgae in pilot-scale open ponds in the presence of organic carbon substrates has not yet been developed. The problems are heterotrophic bacterial contamination and inefficient conversion of organic carbon. Laboratory investigation was combined with outdoor cultivation to find a culture condition that favors the growth of alga, but inhibits bacteria. A window period for mixotrophic cultivation of the alga Graesiella sp. WBG-1 was identified. Using this period, a new sequential phototrophic–mixotrophic cultivation (SPMC) method that enhances algal biomass productivity and limits bacteria contamination at the same time was established for microalgae cultivation in open raceway ponds. Graesiella sp. WBG-1 maximally produced 12.5 g biomass and 4.1 g lipids m−2 day−1 in SPMC in a 1000 m2 raceway pond, which was an over 50% increase compared to phototrophic cultivation. The bacterial number in SPMC (2.97 × 105 CFU ml−1) is comparable to that of the phototrophic cultivations. SPMC is an effective and feasible method to cultivate lipid-rich microalgae in open raceway ponds. Successful scale-up of SPMC in a commercial raceway pond (1000 m2 culture area) was demonstrated for the first time. This method is attractive for global producers of not only lipid-rich microalgae biomass, but also astaxanthin and β-carotene.

The aim of this study was to improve the protein content and yield of heterotrophic microalgal cultivation and establish a simple, economical, and efficient method for microalgal protein production using the novel green alga, Graesiella emersonii WBG-1, which has not been previously reported for heterotrophic cultivation. Through batch heterotrophic cultivation of this alga, we observed that glucose was the optimal carbon source, while it could not use sucrose as a carbon source. Biomass production and protein content were significantly reduced when sodium acetate was used as the carbon source. Compared with nitrate, protein content increased by 93% when urea was used as the nitrogen source. Cultivation temperature had a significant impact on biomass production and protein content. The optimal conditions were glucose as the carbon source at an initial concentration of 10 g/L, urea as the nitrogen source at an initial concentration of 1.62 g/L, and a culture temperature of 35 °C. On the second day of batch cultivation, the highest protein content (66.14%) was achieved, which was significantly higher than that reported in heterotrophic cultures of Chlorella and much higher than that reported for specially established technologies aimed at increasing the protein content, such as two-stage heterotrophic, heterotrophy–dilution–photoinduction, and mixotrophic processes. These results demonstrate the great potential of the heterotrophic cultivation of G. emersonii WBG-1 for protein production.

To understand the effects of physicochemical factors on nitrite transformation by microalgae, a lipid-rich Chlorella with high nitrite tolerance was cultured with 8 mmol/l sodium nitrite as sole nitrogen source under different conditions. The results showed that nitrite transformation was mainly dependent on the metabolic activities of algal cells rather than oxidation of nitrite by dissolved oxygen. Light intensity, temperature, pH, NaHCO3 concentrations, and initial cell densities had significant effects on the rate of nitrite transformation. Single-factor experiments revealed that the optimum conditions for nitrite transformation were light intensity: 300 μmol/m(2); temperature: 30°C; pH: 7-8; NaHCO3 concentration: 2.0 g/l; and initial cell density: 0.15 g/l; and the highest nitrite transformation rate of 1.36 mmol/l/d was achieved. There was a positive correlation between nitrite transformation rate and the growth of Chlorella. The relationship between nitrite transformation rate (mg/l/d) and biomass productivity (g/l/d) could be described by the regression equation y = 61.3x (R(2) = 0.9665), meaning that 61.3 mg N element was assimilated by 1.0 g dry biomass on average, which indicated that the nitrite transformation is a process of consuming nitrite as nitrogen source by Chlorella. The results demonstrated that the Chlorella suspension was able to assimilate nitrite efficiently, which implied the feasibility of using flue gas for mass production of Chlorella without preliminary removal of NOX.