María Cuaresma

Marine Carotenoids: Biological Functions and Commercial Applications

Carotenoids are the most common pigments in nature and are synthesized by all photosynthetic organisms and fungi. Carotenoids are considered key molecules for life. Light capture, photosynthesis photoprotection, excess light dissipation and quenching of singlet oxygen are among key biological functions of carotenoids relevant for life on earth. Biological properties of carotenoids allow for a wide range of commercial applications. Indeed, recent interest in the carotenoids has been mainly for their nutraceutical properties. A large number of scientific studies have confirmed the benefits of carotenoids to health and their use for this purpose is growing rapidly. In addition, carotenoids have traditionally been used in food and animal feed for their color properties. Carotenoids are also known to improve consumer perception of quality; an example is the addition of carotenoids to fish feed to impart color to farmed salmon.

Productivity of Chlorella sorokiniana in a short light-path (SLP) panel photobioreactor under high irradiance

Maximal productivity of a 14 mm light‐path panel photobioreactor under high irradiance was determined. Under continuous illumination of 2,100 µmol photons m−2 s−1 with red light emitting diodes (LEDs) the effect of dilution rate on photobioreactor productivity was studied. The light intensity used in this work is similar to the maximal irradiance on a horizontal surface at latitudes lower than 37°. Chlorella sorokiniana, a fast‐growing green microalga, was used as a reference strain in this study. The dilution rate was varied from 0.06 to 0.26 h−1. The maximal productivity was reached at a dilution rate of 0.24 h−1, with a value of 7.7 g dw m−2 h−1 (m2 of illuminated photobioreactor surface) and a volumetric productivity of 0.5 g dw L−1 h−1. At this dilution rate the biomass concentration inside the reactor was 2.1 g L−1 and the photosynthetic efficiency was 1.0 g dw mol photons. This biomass yield on light energy is high but still lower than the theoretical maximal yield of 1.8 g mol photons−1 which must be related to photosaturation and thermal dissipation of absorbed light energy. Biotechnol. Bioeng. 2009; 104: 352–359

Horizontal or vertical photobioreactors? How to improve microalgae photosynthetic efficiency

The productivity of a vertical outdoor photobioreactor was quantitatively assessed and compared to a horizontal reactor. Daily light cycles in southern Spain were simulated and applied to grow the microalgae Chlorella sorokiniana in a flat panel photobioreactor. The maximal irradiance around noon differs from 400 μmol photons m(-2) s(-1) in the vertical position to 1800 μmol photons m(-2) s(-1) in the horizontal position. The highest volumetric productivity was achieved in the simulated horizontal position, 4 g kg culture(-1) d(-1). The highest photosynthetic efficiency was found for the vertical simulation, 1.3g of biomass produced per mol of PAR photons supplied, which compares favorably to the horizontal position (0.85 g mol(-1)) and to the theoretical maximal yield (1.8 g mol(-1)). These results prove that productivity per unit of ground area could be greatly enhanced by placing the photobioreactors vertically.

Biofilm growth of Chlorella sorokiniana in a rotating biological contactor based photobioreactor

Microalgae biofilms could be used as a production platform for microalgae biomass. In this study, a photobioreactor design based on a rotating biological contactor (RBC) was used as a production platform for microalgae biomass cultivated in biofilm. In the photobioreactor, referred to as Algadisk, microalgae grow in biofilm on vertical rotating disks partially submerged in a growth medium. The objective is to evaluate the potential of the Algadisk photobioreactor with respect to the effects of disk roughness, disk rotation speed and CO2 concentration. These objectives where evaluated in relationship to productivity, photosynthetic efficiency, and long‐term cultivation stability in a lab‐scale Algadisk system. Although the lab‐scale Algadisk system is used, operation parameters evaluated are relevant for scale‐up. Chlorella Sorokiniana was used as model microalgae. In the lab‐scale Algadisk reactor, productivity of 20.1 ± 0.7 g per m2 disk surface per day and a biomass yield on light of 0.9 ± 0.04 g dry weight biomass per mol photons were obtained. Different disk rotation speeds did demonstrate minimal effects on biofilm growth and on the diffusion of substrate into the biofilm. CO2 limitation, however, drastically reduced productivity to 2–4 g per m2 disk surface per day. Productivity could be maintained over a period of 21 weeks without re‐inoculation of the Algadisk. Productivity decreased under extreme conditions such as pH 9–10, temperature above 40°C, and with low CO2 concentrations. Maximal productivity, however, was promptly recovered when optimal cultivation conditions were reinstated. These results exhibit an apparent opportunity to employ the Algadisk photobioreactor at large scale for microalgae biomass production if diffusion does not limit the CO2 supply. Biotechnol. Bioeng. 2014;111: 2436–2445.

Luminostat operation: A tool to maximize microalgae photosynthetic efficiency in photobioreactors during the daily light cycle?

The luminostat regime has been proposed as a way to maximize light absorption and thus to increase the microalgae photosynthetic efficiency within photobioreactors. In this study, simulated outdoor light conditions were applied to a lab-scale photobioreactor in order to evaluate the luminostat control under varying light conditions. The photon flux density leaving the reactor (PFD(out)) was varied from 4 to 20 μmol photons m(-2)s(-1)and the productivity and photosynthetic efficiency of Chlorella sorokiniana were assessed. Maximal volumetric productivity (1.22g kg(-1)d(-1)) and biomass yield on PAR photons (400-700 nm) absorbed (1.27 g mol(-1)) were found when PFD(out) was maintained between 4 and 6 μmol photons m(-2)s(-1). The resultant photosynthetic efficiency was comparable to that already reported in a chemostat-controlled reactor. A strict luminostat regime could not be maintained under varying light conditions. Further modifications to the luminostat control are required before application under outdoor conditions.

Enhanced productivity of a lutein-enriched novel acidophile microalga grown on urea.

Coccomyxa acidophila is an extremophile eukaryotic microalga isolated from the Tinto River mining area in Huelva, Spain. Coccomyxa acidophila accumulates relevant amounts of β-carotene and lutein, well-known carotenoids with many biotechnological applications, especially in food and health-related industries. The acidic culture medium (pH < 2.5) that prevents outdoor cultivation from non-desired microorganism growth is one of the main advantages of acidophile microalgae production. Conversely, acidophile microalgae growth rates are usually very low compared to common microalgae growth rates. In this work, we show that mixotrophic cultivation on urea efficiently enhances growth and productivity of an acidophile microalga up to typical values for common microalgae, therefore approaching acidophile algal production towards suitable conditions for feasible outdoor production. Algal productivity and potential for carotenoid accumulation were analyzed as a function of the nitrogen source supplied. Several nitrogen conditions were assayed: nitrogen starvation, nitrate and/or nitrite, ammonia and urea. Among them, urea clearly led to the best cell growth (~4 × 108 cells/mL at the end of log phase). Ammonium led to the maximum chlorophyll and carotenoid content per volume unit (220 μg·mL·1 and 35 μg·mL·1, respectively). Interestingly, no significant differences in growth rates were found in cultures grown on urea as C and N source, with respect to those cultures grown on nitrate and CO2 as nitrogen and carbon sources (control cultures). Lutein accumulated up to 3.55 mg·g·1 in the mixotrophic cultures grown on urea. In addition, algal growth in a shaded culture revealed the first evidence for an active xanthophylls cycle operative in acidophile microalgae.

Impact of Microalgae-Bacteria Interactions on the Production of Algal Biomass and Associated Compounds

A greater insight on the control of the interactions between microalgae and other microorganisms, particularly bacteria, should be useful for enhancing the efficiency of microalgal biomass production and associated valuable compounds. Little attention has been paid to the controlled utilization of microalgae-bacteria consortia. However, the studies of microalgal-bacterial interactions have revealed a significant impact of the mutualistic or parasitic relationships on algal growth. The algal growth, for instance, has been shown to be enhanced by growth promoting factors produced by bacteria, such as indole-3-acetic acid. Vitamin B12 produced by bacteria in algal cultures and bacterial siderophores are also known to be involved in promoting faster microalgal growth. More interestingly, enhancement in the intracellular levels of carbohydrates, lipids and pigments of microalgae coupled with algal growth stimulation has also been reported. In this sense, massive algal production might occur in the presence of bacteria, and microalgae-bacteria interactions can be beneficial to the massive production of microalgae and algal products. This manuscript reviews the recent knowledge on the impact of the microalgae-bacteria interactions on the production of microalgae and accumulation of valuable compounds, with an emphasis on algal species having application in aquaculture.

Microalgae: Fast-Growth Sustainable Green Factories

Microalgae are definitely on the way to become a sustainable, fast-growing natural green source for highly demanded market products. Research is currently looking at promising outstanding applications of microalgae among which are the production of biofuels from the fatty acids present in biomass, functional foods enriched with microalgae healthy bioactive molecules, and greenhouse gas mitigation (CO2 consuming) based on high cell density microalgae cultures technology. Besides, other traditional microalgae applications are currently increasing in competitiveness in a growing market demand for green biomass, in which microalgae are widely recognized as a healthy, sustainable, and biological renewable resource. Among these traditional applications, microalgae are being used to improve the nutritional quality in animal feed, particularly in aquaculture, therapeutic agent source, and antioxidant activity. In addition, microalgae also show to be functional in waste effluents treatment to remove N and P, thus preventing eutrophication, heavy metals, and others contaminants. This review intends to offer a comprehensive overview of basic aspects of microalgae physiology, massive production of biomass, and current fields of applications at industrial scale.

IDENTIFICATION AND PHYSIOLOGICAL ASPECTS OF A NOVEL CAROTENOID-ENRICHED, METAL-RESISTANT MICROALGA ISOLATED FROM AN ACIDIC RIVER IN HUELVA (SPAIN)1

A heavy‐metal‐resistant, carotenoid‐enriched novel unicellular microalga was isolated from an acidic river in Huelva, Spain. The isolated ribosomal 18S subunit rDNA sequence showed homology with known sequences from green microalgae, the closest sequence (98% homology) belonging to the genus Coccomyxa. The isolated microalga therefore was an up to now uncultured microalga. The microalga was isolated from Tinto River area (Huelva, Spain), an acidic river that exhibits very low pH (1.7–3.1) with high concentrations of sulfuric acid and heavy metals, including Fe, Cu, Mn, Ni, and Al. Electron micrographs show that the microalga contains a large chloroplast with a presence of lipid droplets, an increased number of starch bodies as well as electron‐dense deposits and plastoglobules, the last observed only in iron‐exposed cells. Unlike other acidophile microalgae, the isolated microalga showed high growth rates when cultivated photoautotrophycally (up to 0.6 d−1) in a suitable culture medium prepared at our laboratory. The growth was shown to be iron dependent. When the microalga is grown in fluidized bed reactors, the high growth rates resulted in unexpectedly high productivities for being a microalga that naturally grows in acidic environments (0.32 g·L−1·d−1). The microalga also grows optimally on reduced carbon sources, including glucose and urea, and at an optimal temperature of 35°C. The alga pigment profile is particularly rich in carotenoids, especially lutein, suggesting that the microalga might have potential for antioxidant production, namely, xanthophylls.

Circadian rhythms in the cell cycle and biomass composition of Neochloris oleoabundans under nitrogen limitation.

The circadian clock schedules processes in microalgae cells at suitable times in the day/night cycle. To gain knowledge about these biological time schedules, Neochloris oleoabundans was grown under constant light conditions and nitrogen limitation. Under these constant conditions, the only variable was the circadian clock. The results were compared to previous work done under nitrogen-replete conditions, in order to determine the effect of N-limitation on circadian rhythms in the cell cycle and biomass composition of N. oleoabundans. The circadian clock was not affected by nitrogen-limitation, and cell division was timed in the natural night, despite of constant light conditions. However, because of nitrogen-limitation, not the entire population was able to divide every day. Two subpopulations were observed, which divided alternately every other day. This caused oscillations in biomass yield and composition. Starch and total fatty acids (TFA) were accumulated during the day. Also, fatty acid composition changed during the cell cycle. Neutral lipids were built up during the day, especially in cells that were arrested in their cell cycle (G2 and G3). These findings give insight in the influence of circadian rhythms on the cell cycle and biomass composition.