José María Fernández Sevilla

Protein measurements of microalgal and cyanobacterial biomass

The protein content of dry biomass of the microalgae Porphyridium cruentum, Scenedesmus almeriensis, and Muriellopsis sp. and of the cyanobacteria Synechocystis aquatilis and Arthrospira platensis was measured by the Lowry method following disruption of the cells by milling with inert ceramic particles. The measurements were compared with the Kjeldahl method and by elemental analysis. The nitrogen-to-protein conversion factors for biomass obtained from exponentially growing cells with a steady state doubling time of approximately 23 h were 5.95 for nitrogen measured by Kjeldahl and 4.44 for total nitrogen measured by elemental analysis. The protein content in dry biomass ranged from 30% to 55%. The above conversion factors are useful for estimating the protein content of microalgal biomass produced in rapid steady state growth as encountered in many commercial production processes.

Microalgae, Mass Culture Methods

Introduction Bioreactors for Microalgae Mass Culture Open Raceways Enclosed Photobioreactors Physical Factors That Influence Biomass Productivity Light Availability inside a Photobioreactor The Light Saturation Constant and the Photoinhibition Phenomenon Biomass Productivity Fluid Dynamics and Mixing Gas–Liquid Mass Transfer Temperature Photosynthetic Efficiency Absorbed Photon Flux Operational Considerations Concluding Remarks Nomenclature Acknowledgments Bibliography Keywords: fluid dynamics; gas–liquid mass transfer; light distribution; microalgae mass culture; outdoor production; photosynthetic efficiency; tubular reactor

Assessment of the production of 13C labeled compounds from phototrophic microalgae at laboratory scale

An integrated process for the indoor production of 13C labeled polyunsaturated fatty acids (PUFAs) from Phaeodactylum tricornutum is presented. The core of the process is a bubble column photobioreactor operating with recirculation of the exhaust gas using a low-pressure compressor. Oxygen accumulation in the system is avoided by bubbling the exhaust gas from the reactor in a sodium sulfite solution before returning to it. To achieve a high 13C enrichment in the biomass obtained, the culture medium is initially stripped of carbon, and labeled 13CO(2) is automatically injected on-demand during operation for pH control and carbon supply. The reactor was operated in both batch and semicontinuous modes. In semicontinuous mode, the reactor was operated at a dilution rate of 0.01 h(-1), resulting in a biomass productivity of 0.1 g l(-1) per day. The elemental analysis of the inlet and outlet flows of the reactor showed that 64.9% of carbon was turned into microalgal biomass, 34.9% remained in the supernatant mainly as inorganic compounds. Only 3.8% of injected carbon was effectively fixed as the target labeled product (EPA). Regarding the isotopic composition of fatty acids, results showed that fatty acids were not labeled in the same proportion, the higher the number of carbons the lower the percentage of 13C. Isotopic composition of EPA ranged from 36.5 to 53.5%, as a function of the methodology used (GC-MS, EA-IRMS or gas chromatography-combustion-isotope ratio mass spectrometry (GC-IRMS)). The low carbon uptake efficiency combined with the high cost of 13CO(2) make necessary to redefine the designed culture system to increase the efficiency of the conversion of 13CO(2) into the target product. Therefore, the possibility of removing 12C from the fresh medium, and recovering and recirculating the inorganic carbon in the supernatant and the organic carbon from the EPA depleted biomass was studied. The inorganic carbon of the fresh medium was removed by acidification and stripping with N(2). The inorganic carbon of the supernatant was recovered also by acidification and subsequent stripping with N(2). The operating conditions of this step were optimized for gas flow rate and type of contactor. A carbon recovery step for the depleted biomass was designed based on the catalytic oxidation to CO(2) using CuO (10 wt.%) as catalyst with an oxygen enriched atmosphere (80% O(2) partial pressure). In this way, the carbon losses reduced an 80.2% and the efficiency of the conversion of carbon in EPA was increased to 19.5%, which is close to the theoretical maximum. Further increase in 13CO(2) use efficiency is only possible by additionally recovering other labeled by-products present in the biomass: proteins, carbohydrates, lipids, and pigments.

Production of 13C polyunsaturated fatty acids from the microalga Phaeodactylum tricornutum

An integrated process for the indoor production of 13C labelled PUFA from Phaeodactylum tricornutum is presented. The core of the process is a bubble column photobioreactor from which the exhaust gas from the reactor is returned to the culture by a low pressure compressor. To avoid accumulation of dissolved oxygen in the culture medium, the exhaust gas is bubbled through a sodium sulphite solution before returning it to the reactor. Carbon is removed from the medium before inoculating the alga, then labelled 13CO2 is injected for pH control and carbon supply. The reactor has been operated in semicontinuous mode at a dilution rate of 0.01 h−1, a biomass productivity of 0.1 g L−1 d−1 being obtained. Under this conditions both pH and dissolved oxygen were correctly controlled and the adequacy of the system for autotrophic production of labelled biomass was demonstrated. Analysis by GC-MS revealed that the fatty acids content of the biomass obtained was 10% d.wt., the content of eicosapentaenoic acid was 2.5% d.wt. All the fatty acids were labelled, more that 90% of the carbon present in these fatty acids was 13C. Element analysis of biomass and supernatant showed that 59.5% of injected carbon was assimilated into the biomass whereas 33% remained in the supernatant, and 7.5% remained undetected. Due to the high cost of 13CO2 different strategies for the optimisation of labelled carbon use are proposed.

Effect of pretreatments on biogas production from microalgae biomass grown in pig manure treatment plants

Methane production from pretreated and raw mixed microalgae biomass grown in pig manure was evaluated. Acid and basic pretreatments provided the highest volatile solids solubilisation (up to 81%) followed by alkaline-peroxide and ultrasounds (23%). Bead milling and steam explosion remarkably increased the methane production rate, although the highest yield (377 mL CH4/g SV) was achieved by alkali pretreatment. Nevertheless, some pretreatments inhibited biogas production and resulted in lag phases of 7-9 days. Hence, experiments using only the pretreated solid phase were performed, which resulted in a decrease in the lag phase to 2-3 days for the alkali pretreatment and slightly increased biomass biodegradability of few samples. The limiting step during the BMP test (hydrolysis or microbial inhibition) for each pretreatment was elucidated using the goodness of fitting to a first order or a Gompertz model. Finally, the use of digestate as biofertilizer was evaluated applying a biorefinery concept.

Microalgae: The Basis of Mankind Sustainability

Microalgae were the basis of life into the planet, but only recently these microorganisms are exploited at a commercial scale. Thus, the production of pharmaceuticals, cosmetics, feed, and foods from microalgae is today a commercial reality increasing year by year. Additionally, microalgae have been proposed to be used to enhance the sustainability of existing industrial activities, as wastewater treatment and biofuel production. In this way, the utilization of microalgae at a large scale is considered a green revolution in the sustainability of mankind. This chapter is focused on reviewing the real contribution of microalgae to human activities. The last improvements of technologies and its uses, in addition to still existing bottlenecks for the massive exploitation of these microorgan‐ isms, are reviewed.