Energy crises, global warming, and climatic changes call for technological and commercial advances in manufacturing high-quality transportation fuels from unconventional feedstocks. Microalgae is one of the most promising sources of biofuels due to the high yields attained per unit area and because it does not displace food crops. Neochloris oleabundans (Neo) microalga is an important promising microbial source of single-cell oil (SCO). Different experimental growth and lipid production conditions were evaluated and compared by using optical density (540 nm), dry-weight determination, and flow cytometry (FC). Best Neo average biomass productivity was obtained at 30 degrees C under conditions of nitrogen-sufficiency and CO(2) supplementation (N+/30 degrees C/CO(2)), with an average doubling time of 1.4 days. The second and third highest productivities occurred with N-sufficient cultures without CO(2) supplementation at 26 degrees C (N+/26 degrees C) and at 30 degrees C (N+/30 degrees C), with doubling times of 1.7 and 2.2 days, respectively. Microbial lipid production was monitored by flow cytometry using Nile red (NR), a lipophilic fluorochrome that possesses several advantageous characteristics for in situ screening near real time (at line). Results showed maximum lipid content (56%) after 6 days of nitrogen depletion under nitrogen starvation without CO(2) supplementation (N-/30 degrees C), followed by N-/30 degrees C/CO(2) and N-/26 degrees C conditions with 52% lipid content, after 5 and 6 days of N starvation, respectively. The adequate fatty acid profile and iodine value of Neo lipids reinforced this microalga as a good source of SCO, in particular for use as biodiesel.
Industrial production of novel microalgal isolates is key to improving the current portfolio of available strains that are able to grow in large-scale production systems for different biotechnological applications, including carbon mitigation. In this context, Tetraselmis sp. CTP4 was successfully scaled up from an agar plate to 35- and 100-m3 industrial scale tubular photobioreactors (PBR). Growth was performed semi-continuously for 60 days in the autumn-winter season (17th October – 14th December). Optimisation of tubular PBR operations showed that improved productivities were obtained at a culture velocity of 0.65–1.35 m s−1 and a pH set-point for CO2 injection of 8.0. Highest volumetric (0.08 ± 0.01 g L−1 d−1) and areal (20.3 ± 3.2 g m−2 d−1) biomass productivities were attained in the 100-m3 PBR compared to those of the 35-m3 PBR (0.05 ± 0.02 g L−1 d−1 and 13.5 ± 4.3 g m−2 d−1, respectively). Lipid contents were similar in both PBRs (9–10% of ash free dry weight). CO2 sequestration was followed in the 100-m3 PBR, revealing a mean CO2 mitigation efficiency of 65% and a biomass to carbon ratio of 1.80. Tetraselmis sp. CTP4 is thus a robust candidate for industrial-scale production with promising biomass productivities and photosynthetic efficiencies up to 3.5% of total solar irradiance.
Industrial scale-up of microalgal cultures is often a protracted step prone to culture collapse and the occurrence of unwanted contaminants. To solve this problem, a two-stage scale-up process was developed – heterotrophically Chlorella vulgaris cells grown in fermenters (1st stage) were used to directly inoculate an outdoor industrial autotrophic microalgal production unit (2nd stage). A preliminary pilot-scale trial revealed that C. vulgaris cells grown heterotrophically adapted readily to outdoor autotrophic growth conditions (1-m3 photobioreactors) without any measurable difference as compared to conventional autotrophic inocula. Biomass concentration of 174.5 g L−1, the highest value ever reported for this microalga, was achieved in a 5-L fermenter during scale-up using the heterotrophic route. Inocula grown in 0.2- and 5-m3 industrial fermenters with mean productivity of 27.54 ± 5.07 and 31.86 ± 2.87 g L−1 d−1, respectively, were later used to seed several outdoor 100-m3 tubular photobioreactors. Overall, all photobioreactor cultures seeded from the heterotrophic route reached standard protein and chlorophyll contents of 52.18 ± 1.30% of DW and 23.98 ± 1.57 mg g−1 DW, respectively. In addition to providing reproducible, high-quality inocula, this two-stage approach led to a 5-fold and 12-fold decrease in scale-up time and occupancy area used for industrial scale-up, respectively.
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