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Algae are a promising feedstock for the sustainable production of feed, fuels, and chemicals. Especially in arid regions such as the Arabian Peninsula, algae could play a significant role in enhancing food security, economic diversification, and decarbonization. Within this context, the regional potential of algae commercialization is discussed, exploring opportunities and challenges across technical, societal, and political aspects. Climate, availability of process inputs, and funding opportunities are identified as essential strengths that increase the global competitiveness of regional algae production. Implementation challenges include climate change, securing human resources, and the vital transitioning from research to commercial scales. With balanced management, however, the region's efforts could be the push that is necessary for algal technologies to take off globally.

Multiple models have been developed in the field to simulate growth and product accumulation of microalgal cultures. These models heavily depend on the accurate estimation of growth parameters. In this paper growth parameters are presented for three industrially relevant microalgae species: Nannochloropsis sp., Neochloris oleoabundans, and Picochlorum sp. (BPE23). Dedicated growth experiments were done in photobioreactors to determine the maximal biomass yield on light and maintenance rate, while oxygen evolution experiments were performed to estimate the maximal specific growth rate. Picochlorum sp. exhibited the highest specific growth rate of 4.98 ± 0.24 day−1 and the lowest specific maintenance rate of 0.079 day−1, whereas N. oleoabundans showed the highest biomass yield on light of 1.78 gx·molph−1. The measured growth parameters were used in a simple kinetic growth model for verification. When simulating growth under light conditions as found at Bonaire (12 °N, 68° W), Picochlorum sp. displayed the highest areal biomass productivity of 32.2 g.m−2·day−1 and photosynthetic efficiency of 2.8%. The presented growth parameters show to be accurate compared to experimental data and can be used for model calibration by scientists and industrial communities in the field.

Microalgae are considered a suitable production platform for high-value lipids and oleochemicals. Several species including Nannochloropsis oceanica produce large amounts of essential $$\omega$$ ω -3 polyunsaturated fatty acids (PUFAs) which are integral components of food and feed and have been associated with health-promoting effects. N. oceanica can further accumulate high contents of non-polar lipids with chemical properties that render them a potential replacement for plant oils such as palm oil. However, biomass and lipid productivities obtained with microalgae need to be improved to reach commercial feasibility. Genetic engineering can improve biomass and lipid productivities, for instance by increasing carbon flux to lipids. Here, we report the overexpression of glycerol-3-phosphate acyltransferase (GPAT) in N. oceanica during favorable growth conditions as a strategy to increase non-polar lipid content. Transformants overproducing either an endogenous (NoGPAT) or a heterologous (Acutodesmus obliquus GPAT) GPAT enzyme targeted to the endoplasmic reticulum had up to 42% and 51% increased non-polar lipid contents, respectively, compared to the wild type. Biomass productivities of transformant strains were not substantially impaired, resulting in lipid productivities that were increased by up to 37% and 42% for NoGPAT and AoGPAT transformants, respectively. When exposed to nutrient stress, transformants and wild type had similar lipid contents, suggesting that GPAT enzyme exerts strong flux control on lipid synthesis in N. oceanica under favorable growth conditions. NoGPAT transformants further accumulated PUFAs in non-polar lipids, reaching a total of 6.8% PUFAs per biomass, an increase of 24% relative to the wild type. Overall, our results indicate that GPAT is an interesting target for engineering of lipid metabolism in microalgae, in order to improve non-polar lipid and PUFAs accumulation in microalgae.

Microalgae are considered an efficient accumulator and promising source of Se for feed additive purposes. This study aimed at investigating, for the first time, the effect of phosphorus limitation on Se accumulation and uptake efficiency in N. oceanica. A range of phosphorus concentrations (0–2470 µM) were tested in either the presence or absence of sodium selenite (0, 5, 30 µM). Se accumulation was increased up to 16-fold and Se uptake efficiency was increased up to 3.6-fold under phosphorus growth-limiting concentrations. N. oceanica was then cultivated in a 1.8 L flat-panel photobioreactor in batch operation under two phosphorus growth-limiting concentrations (250 and 750 µM) where the accumulation of Se in the microalgal biomass, as well as its presence in the spent medium were analysed. This study is the first to investigate the effect of phosphorus limitation for increasing Se accumulation in microalgae, and to prevent the release of Se in wastewater.

The major bottleneck in commercializing biofuels and other commodities produced by microalgae is the high cost associated with phototrophic cultivation. Improving microalgal productivities could be a solution to this problem. Synthetic biology methods have recently been used to engineer the downstream production pathways in several microalgal strains. However, engineering upstream photosynthetic and carbon fixation metabolism to enhance growth, productivity, and yield has barely been explored in microalgae. We describe strategies to improve the generation of reducing power from light, as well as to improve the assimilation of CO2by either the native Calvin cycle or synthetic alternatives. Overall, we are optimistic that recent technological advances will prompt long-awaited breakthroughs in microalgal research.

The microalga Nannochloropsis oceanica is considered a promising platform for the sustainable production of high-value lipids and biofuel feedstocks. However, current lipid yields of N. oceanica are too low for economic feasibility. Gaining fundamental insights into the lipid metabolism of N. oceanica could open up various possibilities for the optimization of this species through genetic engineering. Therefore, the aim of this study was to discover novel genes associated with an elevated neutral lipid content. We constructed an insertional mutagenesis library of N. oceanica, selected high lipid mutants by five rounds of fluorescence-activated cell sorting, and identified disrupted genes using a novel implementation of a rapid genotyping procedure. One particularly promising mutant (HLM23) was disrupted in a putative APETALA2-like transcription factor gene. HLM23 showed a 40%-increased neutral lipid content, increased photosynthetic performance, and no growth impairment. Furthermore, transcriptome analysis revealed an upregulation of genes related to plastidial fatty acid biosynthesis, glycolysis and the Calvin-Benson-Bassham cycle in HLM23. Insights gained in this work can be used in future genetic engineering strategies for increased lipid productivity of Nannochloropsis.

Microalgae express high protein levels and can be produced in contained cultivation systems with low water requirements and complete fertilizer use. The production potential is 22-44 tons of protein per hectare per year although the current production scale is small. Techno economic analyses have shown good potential for scale-up and cost reduction. Large-scale production of microalgae in the post-fossil era will rely on the capture of carbon dioxide from the air, or sugars from crops. Microalgal amino acid composition matches well with requirements for food and feed, which, in combination with novel biomass pre-treatment steps, will guarantee high-quality microalgal protein. For a broadening of the microalgae species available as single-cell protein, novel food approval is required.

Tisochrysis lutea is an important microalgal species for fucoxanthin and docosahexaenoic acid (DHA) production with an optimum cultivation temperature of approximately 30 ◦C. The aim of the present work was to develop a winter strain with high productivity at 15 ◦C. The response of the original strain to a decrease in temperature from 30 ◦C to 15 ◦C was investigated in continuous turbidostat experiments. This was followed by adaptation for >180 days at 15 ◦C and 2 rounds of sorting for cells with high chlorophyll fluorescence (top 5%) using fluorescence-activated cell sorting (FACS). For the original strain the productivity of biomass, fucoxanthin, and DHA decreased by 92 %, 98 % and 85 % respectively when decreasing the temperature from 30 ◦C to 15 ◦C. In the sorted cold-adapted ‘winter strain’, biomass, fucoxanthin, and DHA productivities were similar to those at 30 ◦C. In addition, the fucoxanthin concentration increased from 1.11 to 4.24 mg g− 1 dry weight and the polar lipid fraction in total fatty acids increased from 21 % to 55 %. The winter strain showed a robust and stable phenotype after one year of cultivation, expanding the outdoor fucoxanthin and lipid production seasons for this species.

Cellular agriculture could represent a more sustainable alternative to current food and nutraceutical production processes. Tisochrysis lutea microalgae represents a rich source of antioxidants and omega-3 fatty acids essential for human health. However, current downstream technologies are limiting its use. The present work investigates mild targeted acoustic treatment of Tisochrysis lutea biomass at different growth stages and acoustic frequencies, intensities and treatment times. Significant differences have been observed in terms of the impact of these variables on the cell disruption and energy requirements. Lower frequencies of 20 kHz required a minimum of 4500 J to disrupt 90% of the cells, while only 1000 J at 1146 kHz. Comparing these results with current industry standards such as bead milling, up to six times less energy use has been identified. These mild biomass processing approaches offer a certain tunability which could suit a wide range of microorganisms with only minor adjustments.

Besides being considered pathogens, viruses are important drivers of evolution and they can shape large ecological and biogeochemical processes, by influencing host fitness, population dynamics, and community structures. Moreover, they are simple systems that can be used and manipulated to be beneficial and useful for biotechnological applications. In this context, microalgae biotechnology is a growing field of research, which investigated the usage of photosynthetic microorganisms for the sustainable production of food, fuel, chemical, and pharmaceutical sectors. Viruses infecting microalgae have become important subject of ecological studies related to marine and aquatic environments only four decades ago when virus-like-particles associated with bloom-forming algae were discovered. These first findings have opened new questions on evolution and identity. To date, 63 viruses that infect eukaryotic microalgae have been isolated and cultured. In this short review we briefly summarize what is known about viruses infecting eukaryotic microalgae, and how acknowledging their importance can shape future research focussed not only on marine ecology and evolutionary biology but also on biotechnological applications related to microalgae cell factories.

Microalgae have the potential to become microbial cell factories for lipid production. Their ability to convert sunlight and CO2 into valuable lipid compounds has attracted interest from cosmetic, biofuel, food and feed industries. In order to make microalgae-derived products cost-effective and commercially competitive, enhanced growth rates and lipid productivities are needed, which require optimization of cultivation systems and strain improvement. Advances in genetic tool development and omics technologies have increased our understanding of lipid metabolism, which has opened up possibilities for targeted metabolic engineering. In this review we provide a comprehensive overview on the developments made to genetically engineer microalgal strains over the last 30 years. We focus on the strategies that lead to an increased lipid content and altered fatty acid profile. These include the genetic engineering of the fatty acid synthesis pathway, Kennedy pathway, polyunsaturated fatty acid and triacylglycerol metabolisms and fatty acid catabolism. Moreover, genetic engineering of specific transcription factors, NADPH generation and central carbon metabolism, which lead to increase of lipid accumulation are also reviewed.

The effect of light on biomass and fucoxanthin (Fx) productivities was studied in two microalgae, Tisochrysis lutea and Phaeodactylum tricornutum. High and low biomass concentrations (1.1 and 0.4 g L−1) were tested in outdoor pilot‐scale flat‐panel photobioreactors at semi‐continuous cultivation mode. Fluorescence spectroscopy coupled with chemometric modeling was used to develop prediction models for Fx content and for biomass concentration to be applied for both microalgae species. Prediction models showed high R 2 for cell concentration (.93) and Fx content (.77). Biomass productivity was lower for high biomass concentration than low biomass concentration, for both microalgae (1.1 g L−1: 75.66 and 98.14 mg L−1 d−1, for T. lutea and P. tricornutum, respectively; 0.4 g L−1: 129.9 and 158.47 mg L−1 d−1, T. lutea and P. tricornutum). The same trend was observed in Fx productivity (1.1 g L−1: 1.14 and 1.41 mg L−1 d−1, T. lutea and P. tricornutum; 0.4 g L−1: 2.09 and 1.73 mg L−1 d−1, T. lutea and P. tricornutum). These results show that biomass and Fx productivities can be set by controlling biomass concentration under outdoor conditions and can be predicted using fluorescence spectroscopy. This monitoring tool opens new possibilities for online process control and optimization., Graphical abstract Biomass and fucoxanthin outdoor production and monitoring with brown microalgae

The bioeconomy is a new and essential paradigm for reducing our dependence on natural resources and responding to the environmental threats that the Earth is currently facing. In this regard, microalgae offer almost unlimited possibilities for developing a modern bioeconomy given their metabolic flexibility and high biomass output rates, even when produced under harsh conditions, such as when treating wastewaters or using flue gases. In this article, the microalgal contribution to important economic activities such as the production of food and feed, cosmetics and health-related compounds is reviewed. Moreover, potential contributions of microalgae to emerging sectors are discussed, as in the production of biomaterials, agriculture-related products, biofuels and provision of services such as wastewater treatment and the clean-up of industrial gases. The different microalgal production technologies have also been analyzed to identify the main bottlenecks affecting microalgal use in different applications. Finally, the major challenges facing microalgal biotechnology in enlarging its contribution to the bioeconomy are evaluated, and future trends discussed. info:eu-repo/semantics/publishedVersion

This work aimed to select a Tisochrysis lutea phenotype with higher biomass and fucoxanthin productivities using fluorescence-activated cell sorting (FACS). A novel phenotype was obtained after 2 rounds of selection, based on high-fucoxanthin fluorescence. The resulting phenotype forms cell aggregates, has no flagella, and was stable after 15 months. Optimal temperature (30 ◦C) and light (300 μmol m􀀀 2 s􀀀 1) were obtained at laboratory scale, identical to the original strain. The biomass productivity was higher than the original strain: 1.9× at laboratory scale (0.4 L), and 4.5× under outdoor conditions (190 L). Moreover, compared to the original strain, the productivity of fucoxanthin increased 1.6–3.1× and docosahexaenoic acid 1.5–1.9×. These are the highest ever reported outdoor productivities, obtained with a robust new phenotype from a T. lutea monoculture isolated with FACS without genetic manipulation. The resulting phenotype shows high potential for industrial production.

This work aimed to investigate the accumulation of fucoxanthin and lipids in Tisochrysis lutea during growth (N+) and nitrogen-starvation (N−) and to correlate these products with single-cell emissions using fluorescence-activated cell sorting (FACS). Fucoxanthin content decreased 52.94% from N+ to N− in batch cultivation; increased 40.53% as dilution rate changed from 0.16 to 0.55 d−1 in continuous cultivation. Total lipids (N−) were constant (~250 mg/g), but the abundance of neutral lipids increased from 4.87% to 40.63%. Nile red can stain both polar and neutral lipids. However, in vivo, this differentiation is limited due to an overlapping of signals between 600 and 660 nm, caused by neutral lipids concentrations above 3.48% (W/W). Chlorophyll autofluorescence (720 nm) was reported for the first time as a proxy for fucoxanthin (R2 = 0.90) and polar lipids (R2 = 0.98). FACS can be used in high throughput quantification of pigments and lipids and to select and sort cells with high-fucoxanthin/lipids. Paid Open Access

Mathematical models were developed to predict biomass and hydrocarbon productivities and colony size (ouputs) of Botryococcus braunii showa cultures based on light intensity, temperature and dilution rate (inputs). These models predicted the following maximum values: biomass productivity, 1.3 g L−1 d−1; hydrocarbon productivity, 1.5 mg L−1 d−1; colony size, 320 µm under different culture conditions respectively. These values were confirmed experimentally. Additionally, the combination of inputs that simultaneously maximize all the possible outputs combinations were determined. The prediction for biomass-hydrocarbon-colony size were 1 g L−1 d−1, 12.05 mg L−1 d−1 and 156.8 µm respectively; biomass productivity-hydrocarbon productivities: 1 g L−1 d−1 and 13.94 mg L−1 d−1 respectively; biomass productivity-colony size: 1 g L−1 d−1 and 172.8 µm respectively; hydrocarbon productivity-colony size: 9 mg L−1 d−1 and 240 µm respectively. All these predictions were validated experimentally. These models might be very useful to implement a Botryococcus braunii showa large scale production. publishedVersion

Botryococcus braunii can produce both long-chain hydrocarbons as well as carbohydrates in large quantities, and is therefore a promising industrial organism for the production of biopolymer building blocks. Many studies describe the use of different strains of Botryococcus braunii but differences in handling and cultivation conditions make the comparison between strains difficult. In this study, 16 B. braunii strains obtained from six culture collections were compared for their biomass productivity and hydrocarbon and carbohydrate content. Biomass productivity was highest for AC768 strain with 1.8 g L−1 day−1, while hydrocarbon production ranged from none to up to 42% per gram biomass dry weight, with Showa showing the highest hydrocarbon content followed by AC761. The total carbohydrate content varied from 20% to 76% per gram of the biomass dry weight, with CCALA777 as the highest producer. Glucose and galactose are the main monosaccharides in most strains and fucose content reached 463 mg L−1 in CCALA778.

In this work, the mild disintegration of the microalgae Chlorella vulgaris for the release of intracellular products has been studied. By means of bead milling the microalgae suspensions were successfully disintegrated at different biomass concentrations (25-145 gDW kg(-1)) over a range of agitator speeds (6-12 m s(-1)). In all cases over 97% of cell disintegration was achieved resulting in a release of water soluble proteins. A clear optimum rate of disintegration and protein release was observed at an agitator speed of 9-10 m s(-1) regardless of the biomass concentration. Selective extraction of water soluble proteins was observed as proteins released sooner than cell disintegration took place. Proteins could be released at 85% lower energy input than for cell disintegration resulting in specific energy consumptions well below 2.5 kWh kgDW(-1).

The disintegration of three industry relevant algae (Chlorella vulgaris, Neochloris oleoabundans and Tetraselmis suecica) was studied in a lab scale bead mill at different bead sizes (0.3–1 mm). Cell disintegration, proteins and carbohydrates released into the water phase followed a first order kinetics. The process is selective towards proteins over carbohydrates during early stages of milling. In general, smaller beads led to higher kinetic rates, with a minimum specific energy consumption of ⩽0.47 kWh kgDW−1 for 0.3 mm beads. After analysis of the stress parameters (stress number and stress intensity), it appears that optimal disintegration and energy usage for all strains occurs in the 0.3–0.4 mm range. During the course of bead milling, the native structure of the marker protein Rubisco was retained, confirming the mildness of the disruption process.

Microalgae are a promising source for proteins, lipids and carbohydrates for the food/feed and biofuel industry. In comparison with soya and palm oil, microalgae can be grown in a more efficient and sustainable way. To make microalgae production economically feasible it is necessary to optimally use all produced compounds. To accomplish this focus needs to be put on biorefinery techniques which are mild and effective. Of the techniques described, Pulsed Electric Field (PEF) seems to be the most developed technique compared to other cell disruption applications. For separation technology ionic liquids seems most promising as they are able to both separate hydrophobic and hydrophilic compounds. But additional studies need to be evolved in the coming years to investigate their relevance as novel cell disruption and separation methods. We propose a complete downstream processing flow diagram that is promising in terms of low energy use and state of the art knowledge.

The synergistic effect of temperature (25-65 °C) and total specific energy input (0.55-1.11 kWh kgDW(-1)) by pulsed electric field (PEF) on the release of intracellular components from the microalgae Chlorella vulgaris was studied. The combination of PEF with temperatures from 25 to 55 °C resulted in a conductivity increase of 75% as a result of cell membrane permeabilization. In this range of temperatures, 25-39% carbohydrates and 3-5% proteins release occurred and only for carbohydrate release a synergistic effect was observed at 55 °C. Above 55 °C spontaneous cell lysis occurred without PEF. Combined PEF-temperature treatment does not sufficiently disintegrate the algal cells to release both carbohydrates and proteins at yields comparable to the benchmark bead milling (40-45% protein, 48-58% carbohydrates).

In this feature, leading researchers in the field of microbial biotechnology speculate on the technical and conceptual developments that will drive innovative research and open new vistas over the next few years

Acceleration-stat (A-stat) cultivations in which the dilution rate is continuously changed at a constant acceleration rate, leading to different average light intensities inside the photobioreactor, can supply more information and reduce experimental time compared with chemostat cultivations. The A-stat was used to optimize the biomass and product yield of continuous cultures of the microalgae D. tertiolecta in a flat-panel reactor. In this study, four different accelerations were studied, a pseudo steady state was maintained at acceleration rates of 0.00016 and 0.00029 h-2 and results were similar to those of the chemostat. An increase in the acceleration rate led to an increase in the deviation between results obtained in the A-stat and in the chemostats. We concluded that it is advantageous to use the A-stat instead of chemostats to determine culture characteristics and optimize a specific photobioreactor. The effect of average light intensity inside the photobioreactor on the production of vitamins C and E, lutein, and β-carotene was studied using the A-stat. The highest concentrations of these products were 3.48 ± 0.46, 0.33 ± 0.06, 5.65 ± 0.24, and 2.36 ± 0.38 mg g-1 respectively. These results were obtained at different average light intensities, showing the importance of optimizing each product on light intensity. © 2004 Wiley Periodicals, Inc.

The present work established an efficient staining method for fluorescence activated cell sorting (FACS) with Chlorococcum littorale maintaining cellular viability. The method was designed to detect high-lipid cells and to guarantee cellular viability. BODIPY505/515 (BP) was more suitable to FACS when compared to Nile red. The optimum concentrations were 0.4 μg ml−1 of BP, 0.1% DMSO or 0.35% ethanol. Both ethanol and DMSO were equally efficient and assured cellular viability after the staining and sorting. Here a method is presented to rapidly screen and sort lipid rich cells of C. littorale with FACS, which can be used to produce new inoculum with increased cellular lipid content.

Hydrogen is a clean energy alternative to fossil fuels. Photosynthetic bacteria produce hydrogen from organic compounds by an anaerobic light-dependent electron transfer process. In the present study hydrogen production by three photosynthetic bacterial strains (Rhodopseudomonas sp., Rhodopseudomonas palustris and a non-identified strain), from four different short-chain organic acids (lactate, malate, acetate and butyrate) was investigated. The effect of light intensity on hydrogen production was also studied by supplying two different light intensities, using acetate as the electron donor. Hydrogen production rates and light efficiencies were compared. Rhodopseudomonas sp. produced the highest volume of H2. This strain reached a maximum H2 production rate of 25 ml H2 l(-1) h(-1), under a light intensity of 680 micromol photons m(-2) s(-1), and a maximum light efficiency of 6.2% under a light intensity of 43 micromol photons m(-2) s(-1). Furthermore, a decrease in acetate concentration from 22 to 11 mM resulted in a decrease in the hydrogen evolved from 214 to 27 ml H2 per vessel.

The prospect of sustainable production of food ingredients from photoautotrophic microalgae was reviewed. Clearly, there is scope for microalgal oils to replace functions of major vegetable oils, and in addition to deliver health benefits to food products. Furthermore, with a limited production surface, a substantial portion of the European Union market could be supplied with edible oils and proteins from microalgae. Yet, before microalgal ingredients can become genuinely sustainable and cost effective alternatives for current food commodities, major breakthroughs in production technology and in biorefinery approaches are required. Moreover, before market introduction, evidence on safety of novel microalgal ingredients, is needed. In general, we conclude that microalgae have a great potential as a sustainable feedstock for food commodities.

Worldwide, microalgal biofuel production is being investigated. It is strongly debated which type of production technology is the most adequate. Microalgal biomass production costs were calculated for 3 different micro algal production systems operating at commercial scale today: open ponds, horizontal tubular photobioreactors and flat panel photobioreactors. For the 3 systems, resulting biomass production costs including dewatering, were 4.95, 4.15 and 5.96 € per kg, respectively. The important cost factors are irradiation conditions, mixing, photosynthetic efficiency of systems, medium- and carbon dioxide costs. Optimizing production with respect to these factors, a price of € 0.68 per kg resulted. At this cost level microalgae become a promising feedstock for biodiesel and bulk chemicals.Photobioreactors may become attractive for microalgal biofuel production.

The feasibility of microalgae production for biodiesel was discussed. Although algae are not yet produced at large scale for bulk applications, there are opportunities to develop this process in a sustainable way. It remains unlikely, however, that the process will be developed for biodiesel as the only end product from microalgae. In order to develop a more sustainable and economically feasible process, all biomass components (e.g. proteins, lipids, car- bohydrates) should be used and therefore biorefi ning of microalgae is very important for the selective separation and use of the functional biomass components. If biorefi ning of microalgae is applied, lipids should be fractionated into lipids for biodiesel, lipids as a feedstock for the chemical industry and w -3 fatty acids, proteins and carbohydrates for food, feed and bulk chemicals, and the oxygen produced should be recovered also. If, in addition, production of algae is done on residual nutrient feedstocks and CO2, and production of microalgae is done on a large scale against low production costs, production of bulk chemicals and fuels from microalgae will become economically feasible. In order to obtain that, a number of bottlenecks need to be removed and a multidisciplinary approach in which sys- tems biology, metabolic modeling, strain development, photobioreactor design and operation, scale-up, biorefi ning, integrated production chain, and the whole system design (including logistics) should be addressed. © 2010 Society of Chemical Industry and John Wiley & Sons, Ltd

In the present work we identified and quantified the effect of hydrodynamic stress on two different microalgae strains, Dunaliella tertiolecta and D. salina, cultivated in bench-scale bubble columns. The cell death rate constant increased with increasing gas-entrance velocity at the sparger. Dunaliella salina was slightly more sensitive than D. tertiolecta. The critical gas-entrance velocities were approximately 50 and 30 m s(-1) for D. tertiolecta and D. salina, respectively. The effects of gas-flow rate, culture height, and nozzle diameter on the death rate constant were also studied. From these results it was concluded that bubble rising and bubble bursting are not responsible for cell death. Regarding nozzle diameter, small nozzles were more detrimental to cells. The bubble formation at the sparger was found to be the main event leading to cell death.

The slow development of microalgal biotechnology stems from the failure in the design of large-scale photobioreactors where light energy is efficiently utilized. Due to the light gradient inside the reactor and depending on the mixing properties, algae are subjected to certain light/dark cycles where the light period is characterized by a light gradient. These light/dark cycles will determine productivity and biomass yield on light energy. Air-lift reactors can be used for microalgae cultivation and medium-frequency light/dark cycles will be found in these systems. Light/dark cycles are associated with two basic parameters: first, the light fraction, i.e., the ratio between the light period and the cycle time and second, the frequency of the light/dark cycle. In the present work, light/dark cycles found in air-lift reactors were simulated taking into account the light gradient during the light period. The effect of medium-frequency cycle time (10-100 s) and light fraction (0.1-1) on growth rate and biomass yield on light energy of the microalgae Dunaliella tertiolecta was studied. The biomass yield and growth rates were mainly affected by the light fraction, while cycle time had little influence. Response surface methodology was used and a statistical model describing the effect of light fraction and cycle time on growth rate and biomass yield on light energy was developed. The use of the model as a reactor design criterion is discussed.

High-quality research in the field of marine biotechnology is one of the key-factors for successful innovation in exploiting the vast diversity of marine life. However, fascinating scientific research with promising results and claims on promising potential applications (e.g. for pharmaceuticals, nutritional supplements, (feed-)products for aquaculture and bioremediation solutions) is not the only factor to realise the commercial applications of marine biotechnology. What else is needed to exploit the promising potential of marine biotechnology and to create new industrial possibilities? In the study project 'Ocean Farming-Sustainable exploitation of marine organisms', we explore the possibilities of marine organisms to fulfill needs, such as safe and healthy food, industrial (raw) materials and renewable energy in a sustainable way. One of the three design groups is envisioning the future of strong land-based 'marine' market chains. Marine biotechnology is one of the foci of attention in this design group. This article provides a model of future-oriented thinking in which a variety of experts actively participate. (C) 2003 Elsevier B.V. All rights reserved.

The effect of high superficial gas velocities in continuous and batch cultures of the strains Dunaliella tertiolecta, Chlamydomonas reinhardtii wild-type and cell wall-lacking mutant was studied in bubble columns. No cell damage was found for D. tertiolecta and C. reinhardtii (wild-type) up to superficial gas velocities of 0.076 and 0.085 m s(-1), respectively, suggesting that high superficial gas velocities alone cannot be responsible for cell death and, consequently, bubble bursting cannot be the sole cause for cell injury. A death rate of 0.46 +/- 0.08 h(-1) was found for C. reinhardtii (cell wall-lacking mutant) at a superficial gas velocity of 0.076 m s(-1), and increased to 1.01 +/- 0.29 h(-1) on increasing superficial gas velocity to 0.085 m s(-1). Shear sensitivity is thus strain-dependent and to some extent the cell wall plays a role in the protection against hydrodynamic shear. When studying the effect of bubble formation at the sparger in batch cultures of D. tertiolecta by varying the number of nozzles, a death rate of 0.047 +/- 0.016 h(-1) was obtained at high gas entrance velocities. D. tertiolecta was cultivated in a pilot-plant reactor under different superficial gas velocities of up to 0.026 m s(-1), with relatively low gas entrance velocities and no cell damage was observed. There is some indication that the main parameter causing cell death and damage was the gas entrance velocity at the sparger.

In the present work we wanted to know what happens during time to biomass and lipid productivities of Chlorococcum littorale repeatedly subjected to N-starvation. Experiments were done using repeated cycles of batch-wise N run-out (after 2 days N = 0). Two different cycles were used: repeated short-starvation (6 days of N = 0) over a total period of 72 days and repeated long-starvation (13 days of N = 0) over a total period of 75 days. Batches (using fresh inocula) were done separately as control. Shorter and longer periods of starvation showed no differences in biomass productivities and PSII quantum yield evolution. The repeated short-starvation-batches showed the same lipid productivities as the control short-starvation batches. Most importantly, the biomass lipid content was the same between control and repeated-batches. Altogether, the results point to C. littorale as a resilient and stable strain, with potential to be used under semi continuous cultivation.

To optimize fucoxanthin production inTisochrysis lutea, the effect of different process parameters on fucoxanthin productivity (Pfx) were evaluated using batch and continuous experiments. In batch, the highest Pfx was found at 30°C and 300μmolm−2s−1, allowing to design continuous experiments to optimize the dilution rate. The highest ever reported Pfx (9.43–9.81mgL−1d−1) was achieved at dilution rates of 0.53 and 0.80d−1. Irradiance was varied (50–500μmolm−2s−1) to result in a range of absorbed light between 2.23 and 25.80molm−2d−1at a fixed dilution rate (0.53d−1). These experiments validated the hypothesis that light absorbed can be used to predict fucoxanthin content, resulting in 2.23molm−2d−1triggering the highest fucoxanthin content (16.39mg/g). The highest Pfx was found with 18.38molm−2d−1. These results can be used to achieve high Pfx or fucoxanthin content during cultivation ofTisochrysis lutea.