Quantification of oxygen production and respiration rates in mixotrophic cultivation of microalgae in nonstirred photobioreactors

Online monitoring and controlling of different cellular parameters are key issues in aerobic bioprocesses. Since mixotrophic cultivation, in which we observe a mixture of cellular respiration and oxygen production has gained more popularity, there is a need for an on‐process quantification of these parameters. The presented and adapted double gassing‐out method applied to a mixotrophic cultivation of Galdieria sulphuraria , will be a tool for monitoring and further optimization of algal fermentation in nonstirred photobioreactors (PBR). We measured the highest net specific oxygen production rate (opr _net) as 5.73 · 10^?3 mol_O2 g^?1 h^?1 at the lowest oxygen uptake rate (OUR) of 1.00 · 10^?4 mol_O2 L^?1 h^?1. Due to higher cell densities, we also demonstrated the increasing shading effect by a decrease of opr _net, reaching the lowest value of 1.25 10^?5 mol_O2 g^?1 h^?1. Nevertheless, with this on process measurement, we can predict the relation between the zone in which oxygen is net produced to the area where cell respiration dominates in a PBR, which has a major impact to optimize cell growth along with the formation of different products of interest such as pigments.     

Is axenicity crucial to cryopreserve microalgae?

Large culture collections of microalgae and cyanobacteria such as the Coimbra Collection of Algae (ACOI) hold unialgal cultures consisting of a population of cells/colonies of a certain species. These cultures are usually non-axenic, as other organisms such as bacteria and microfungi are also present in culture due to co-isolation. Attention has been recently given to partner organisms since studies indicate that some bacteria are important for nutrient uptake of the algal cells, acting as simbionts. Despite this benign effect in the actively growing cultures, when cryopreservation is applied for inactive-stage storage, these organisms may recover faster than the algae, thus affecting their recovery and the viability assessments. In this study, a set of mucilaginous ACOI microalgae were selected, cell features known for their relevance in cryopreservation success were recorded and simple two-step cryopreservation tests were applied. Thawed samples were transferred to fresh culture medium for recovery. Viability was assessed and partner organism proliferation (pop) was recorded. Results were analyzed by t-tests. Statistical models allowed us to support the known tendency for small, unicellular algae with no outer structures to be successfully cryopreserved and the negative effect of vacuoles in the cell prior to cryopreservation. On average cryopreservation with MeOH or Me₂SO led to the recovery of nearly half the cells. It was found that the cryoprotection step with MeOH is when pop is triggered and that the use of Me₂SO can prevent this effect. Progress on understanding the cultured consortia will assist the improvement of cryopreservation and research using microalgal cultures.     

Recovery of microalgal biomass and metabolites: process options and economics

Commercial production of intracellular microalgal metabolites requires the following: (1) large-scale monoseptic production of the appropriate microalgal biomass; (2) recovery of the biomass from a relatively dilute broth; (3) extraction of the metabolite from the biomass; and (4) purification of the crude extract. This review examines the options available for recovery of the biomass and the intracellular metabolites from the biomass. Economics of monoseptic production of microalgae in photobioreactors and the downstream recovery of metabolites are discussed using eicosapentaenoic acid (EPA) recovery as a representative case study.     

The potential of sustainable algal biofuel production using wastewater resources

The potential of microalgae as a source of renewable energy has received considerable interest, but if microalgal biofuel production is to be economically viable and sustainable, further optimization of mass culture conditions are needed. Wastewaters derived from municipal, agricultural and industrial activities potentially provide cost-effective and sustainable means of algal growth for biofuels. In addition, there is also potential for combining wastewater treatment by algae, such as nutrient removal, with biofuel production. Here we will review the current research on this topic and discuss the potential benefits and limitations of using wastewaters as resources for cost-effective microalgal biofuel production.     

Life-cycle analysis on biodiesel production from microalgae: Water footprint and nutrients balance

This research examines the life-cycle water and nutrients usage of microalgae-based biodiesel production. The influence of water types, operation with and without recycling, algal species, geographic distributions are analyzed. The results confirm the competitiveness of microalgae-based biofuels and highlight the necessity of recycling harvested water and using sea/wastewater as water source. To generate 1 kg biodiesel, 3726 kg water, 0.33 kg nitrogen, and 0.71 kg phosphate are required if freshwater used without recycling. Recycling harvest water reduces the water and nutrients usage by 84% and 55%. Using sea/wastewater decreases 90% water requirement and eliminates the need of all the nutrients except phosphate. The variation in microalgae species and geographic distribution are analyzed to reflect microalgae biofuel development in the US. The impacts of current federal and state renewable energy programs are also discussed to suggest suitable microalgae biofuel implementation pathways and identify potential bottlenecks.     

Chlorella species as hosts for genetic engineering and expression of heterologous proteins: Progress,challenge and perspective

The species of Chlorella represent a highly specialized group of green microalgae that can produce high levels of protein. Many Chlorella strains can grow rapidly and achieve high cell density under controlled conditions and are thus considered to be promising protein sources. Many advances in the genetic engineering of Chlorella have occurred in recent years, with significant developments in successful expression of heterologous proteins for various applications. Nevertheless, a lot of obstacles remain to be addressed, and a sophisticated and stable Chlorella expression system has yet to emerge. This review provides a brief summary of current knowledge on Chlorella and an overview of recent progress in the genetic engineering of Chlorella, and highlights the advances in the development of a genetic toolbox of Chlorella for heterologous protein expression. Research directions to further exploit the Chlorella expression system with respect to both challenges and perspectives are also discussed. This paper serves as a comprehensive literature review for the Chlorella community and will provide valuable insights into future exploration of Chlorella as a promising host for heterologous protein expression.     

Metabolomics integrated with transcriptomics and proteomics: Evaluation of systems reaction to nitrogen deficiency stress in microalgae

Microalgae have higher productivity of biomass than the conventional crops of fuel and are therefore, considered a potential biofuel source. Lipid, an important precursor of biodiesel, can be overproduced in microalgae by nitrogen deprivation. During nitrogen deficiency, radicals are overproduced, and the antioxidant levels are insufficient to counteract the radicals. Thus, the increase in cellular oxidative stress level, consequently acts as a stimulus for lipid accumulation. Lipid accumulation requires an excess of acetyl CoA and NADPH that is made possible by the following mechanism. Glycolysis upregulation overproduces pyruvate, which could be further transformed into acetyl CoA by the pyruvate dehydrogenase complex; while the upregulation of the oxidative pentose phosphate cycle generates a high amount of NADPH. In addition to lipid overproduction, the lack of nitrogen often causes the accumulation of carbohydrates in selected species of microalgae, which could be used to generate biogas and bioethanol from the defatted biomass. By providing details on the differential regulation of the biochemical pathways leading to lipid and carbohydrate accumulation in nitrogen starved microalgae, the review opens up new possibilities in the microalgal biofuel production.     

Production cost of a real microalgae production plant and strategies to reduce it

The cost analysis of a real facility for the production of high value microalgae biomass is presented. The facility is based on ten 3 m³ tubular photobioreactors operated in continuous mode for 2 years, data of Scenedesmus almeriensis productivity but also of nutrients and power consumption from this facility being used. The yield of the facility was close to maximum expected for the location of Almería, the annual production capacity being 3.8 t/year (90 t/ha·year) and the photosynthetic efficiency being 3.6%. The production cost was 69 €/kg. Economic analysis shows that labor and depreciation are the major factors contributing to this cost. Simplification of the technology and scale-up to a production capacity of 200 t/year allows to reduce the production cost up to 12.6 €/kg. Moreover, to reduce the microalgae production cost to approaches the energy or commodities markets it is necessary to reduce the photobioreactor cost (by simplifying its design or materials used), use waste water and flue gases, and reduce the power consumption and labor required for the production step. It can be concluded that although it has been reported that production of biofuels from microalgae is relatively close to being economically feasible, data here reported demonstrated that to achieve it by using the current production technologies, it is necessary to substantially reduce their costs and to operate them near their optimum values.     

Grazing and UV radiation effects on an Antarctic intertidal microalgal assemblage: a long-term field study

A 15 week field experiment (austral summer Nov–Mar) was carried out in an intertidal hard bottom platform in Antarctica (King George Island). To test whether grazing and ultraviolet radiation (UVR) influenced the succession of a benthic microalgal assemblage, a two-factorial design was used (1) ambient radiation, >280 nm; (2) ambient minus UV-B, >320 nm; (3) ambient minus UVR, >400 nm versus grazer–no grazer). On four sampling occasions microalgae were identified, counted and carbon contents were calculated. The assemblage was dominated by the diatom genera Navicula and Cocconeis. Biomass was generally low in all treatments but was significantly reduced by grazing throughout the experiment. No significant UV effects were found. Grazer absence particularly favoured diatoms of the genus Cocconeis. We conclude that the Antarctic microalgal assemblage was unaffected by present day UVR whereas grazers acted as important drivers on the intertidal microalgal community structure.     

Microalgae-based carbohydrates for biofuel production

Microalgae are considered as the most promising renewable feedstock for biofuel production and biorefineries, due to their advantages of fast growth, efficient carbon dioxide fixation, not competing for arable lands and potable water, and potentially accumulating high amounts of lipids and carbohydrates. Since carbohydrates in microalgae biomass are mainly cellulose in the cell wall and starch in the plastids without lignin and low hemicelluloses contents, they can be readily converted into fermentable sugars. However, to date there are very few studies focusing on the use of microalgae-based carbohydrates for biofuel production, which requires more understanding and knowledge to support the technical feasibility of this next-generation feedstock. This review article elucidates comprehensive information on the characteristics and metabolism of main fermentable microalgal carbohydrates (e.g., starch and cellulose), as well as the key factors and challenges that should be addressed during production and saccharification of microalgal carbohydrates. Furthermore, developments on the utilization of microalgae-based feedstock in producing liquid and gaseous biofuels are summarized. The objective of this article is to provide useful knowledge and information with regard to biochemistry, bioprocess engineering, and commercial applications to assist in the viable technology development of for biofuels generation from microalgae-based carbohydrates.