Trait drift in microalgae and applications for strain improvement.

Microalgae are increasingly used to generate a wide range of commercial products, and there is growing evidence that microalgae-based products can be produced sustainably. However, industrial production of microalgal biomass is not as developed as other biomanufacturing platform technologies. In addition, results of bench-scale research often fail to translate to large-scale or mass production systems. This disconnect may result from trait drift and evolution occurring, through time, in response to unique drivers in each environment, such as cultivation regimes, weather, and pests. Moreover, outdoor and indoor cultivation of microalgae has the potential to impose negative selection pressures, which makes the maintenance of desired traits a challenge. In this context, this review sheds the light on our current understanding of trait drift and evolution in microalgae. We delineate the basics of phenotype plasticity and evolution, with a focus on how microalgae respond under various conditions. In addition, we review techniques that exploit phenotypic plasticity and evolution for strain improvement in view of industrial commercial applications, highlighting associated advantages and shortcomings. Finally, we suggest future research directions and recommendations to overcome unwanted trait drift and evolution in microalgae cultivation.

RICTOR Affects Melanoma Tumorigenesis and Its Resistance to Targeted Therapy.

The network defined by phosphatidylinositol-3-kinase (PI3K), AKT, and mammalian target of rapamycin (mTOR) plays a major role in melanoma oncogenesis and has been implicated in BRAF inhibitor resistance. The central role of RICTOR (rapamycin-insensitive companion of mTOR) in this pathway has only recently begun to be unraveled. In the present study, we assessed the role of mTORC2/RICTOR in BRAF-mutated melanomas and their resistance to BRAF inhibition. We showed that RICTOR was significantly overexpressed in melanoma and associated with bad prognoses. RICTOR overexpression stimulated melanoma-initiating cells (MICs) with 'stemness' properties. We also showed that RICTOR contributed to melanoma resistance to BRAF inhibitors and rendered the cells very sensitive to mTORC2 inhibition. We highlighted a connection between mTORC2/RICTOR and STAT3 in resistant cells and revealed an interaction between RAS and RICTOR in resistant melanoma, which, when disrupted, impeded the proliferation of resistant cells. Therefore, as a key signaling node, RICTOR contributes to BRAF-dependent melanoma development and resistance to therapy and, as such, is a valuable therapeutic target in melanoma.

Effect of Mild Salinity Stress on the Growth, Fatty Acid and Carotenoid Compositions, and Biological Activities of the Thermal Freshwater Microalgae Scenedesmus sp.

Carotenoids have strong antioxidant activity as well as therapeutic value. Their production has been induced in algae under stressful culture conditions. However, the extreme culture conditions lead to the Programmed Cell Death (PCD) of algae, which affects their growth and productivity. This study was performed to evaluate the effect of salinity on the physiological and biochemical traits of Scenedesmus sp., thermal freshwater microalgae from Northern Tunisia. It was cultured under different NaCl concentrations ranging from 0 to 60 g/L. Results showed a good growth and high contents of total chlorophyll and carotenoids in Scenedesmus sp. cultured at 10 g/L of NaCl (salt-stressed 10 (Ss10)). The pigment composition of the Ss10 extract was acquired using HPLC-MS, and showed that the carotenoid fraction is particularly rich in xanthophylls. Moreover, the antioxidant (DPPH and FRAP) and enzymatic inhibition (tyrosinase and elastase) activities of the Ss10 extract were higher compared to those of the control culture. In addition, the cytotoxicity test on B16 cells showed that the Ss10 extract was non-toxic for all tested concentrations below 100 µg/mL. It also showed a rich unsaturated fatty acid (FA) composition. Therefore, these findings suggest that Scenedesmus sp. strain cultivated under mild stress salinity could be a source of biomolecules that have potential applications in the nutraceutical and cosmeceutical industries.

Pilot-scale outdoor production of Scenedesmus sp. in raceways using flue gases and centrate from anaerobic digestion as the sole culture medium.

This work investigated the production of Scenedesmus sp. in semi-continuous mode in three pilot-scale outdoor raceways (7.2 m2) using flue gas for CO2 supply and centrate from the anaerobic digestion of urban wastewater as the sole nutrient source. Experiments were performed at different culture depths, 5, 10 and 15 cm, while evaluating two centrate concentrations (30% and 45%) at dilution rates of 0.2 and 0.3 d-1. Under optimal conditions of 30% centrate, 0.3 d-1 dilution rate and a 15 cm culture depth, a maximum biomass productivity of 22.9 g m-2 d-1 was obtained. The optical properties of the cultures were studied and the results showed a photosynthetic efficiency of up to 2.0% and a quantum yield of 0.3 g biomass E-1. Nitrogen and phosphorus removal rates of 3 g N m-2 d-1 and 0.6 g P m-2 d-1 were recorded, respectively. Lipid productivity of 2.3 g m-2 d-1 was determined possessing a suitable fatty acids profile for biofuel production.

The role of RICTOR downstream of receptor tyrosine kinase in cancers.

The importance of the network defined by phosphatidylinositol-3-kinase (PI3K), AKT and mammalian target of rapamycin (mTOR) downstream of Receptor Tyrosine Kinase (RTK) has been known for many years but the central role of RICTOR (rapamycin-insensitive companion of mTOR) in this pathway is only starting to emerge. RICTOR is critical for mTORC2 (the mammalian target of rapamycin complex 2) kinase activity and as such plays a key role downstream of RTK. Alterations of RICTOR have been identified in a number of cancer cell types and its involvement in tumorigenesis has begun to be unraveled recently. Here, we summarize new research into the biology of RICTOR signaling in cancers focusing on tumors with altered RTK. We show that, as a key signaling node and critical effector of RTKs, RICTOR is becoming a valuable therapeutic target in cancer with altered RTK.

Utilization of centrate from urban wastewater plants for the production of Scenedesmus sp. in a raceway-simulating reactor.

This work investigates the production of the native microalgae strain Scenedesmus sp. in semi-continuous mode at lab scale in open raceway-simulating reactors and using centrate as the culture medium. The biomass productivity and nutrient removal capacity of Scenedesmus sp. at different dilution rates were investigated indoors as well as its tolerance to centrate as the culture medium at different concentrations. A biomass productivity of 7.80 g/m2 day was obtained at 200 µE/m2 s, 5 cm culture depth, 0.30 1/day of dilution rate and 60% centrate while nitrogen and phosphorus removal rates were 1.50 g/m2 day and 0.15 g/m2 day, respectively. The produced biomass characterization under these conditions showed a lipid content of 12.60% d wt. along with a favorable fatty acids profile with 57.70% of total fatty acids composed of saturated and monounsaturated fatty acids. Subsequently, the effect of light intensity and culture depth on biomass productivity and nutrient uptake as well as the biochemical composition and fatty acids profile was studied using two irradiance levels (200 and 1000 µE/m2 s) and four culture depths (5 cm, 10 cm, 15 cm and 20 cm). Under optimal conditions of 1000 µE/m2 s, 60% centrate, 0.30 1/day dilution rate and 15 cm culture depth, a maximum biomass productivity of 22.20 g/m2 day was obtained. Nitrogen and phosphorus removal rates of 2.00 gN/m2 day and 0.40 gP/m2 day, respectively, were recorded. An amount of 11.70% d wt. of lipids was determined along with a suitable fatty acids profile for biofuel production.

Characterization of Amphora sp., a newly isolated diatom wild strain, potentially usable for biodiesel production.

Microalgae as feedstock for biofuel production have attracted serious consideration as an important sustainable source of energy. For biodiesel production with microalgae, a series of consecutive processes should be performed as selection of adequate microalgal strains, mass culture, cell harvesting, oil extraction and transesterification. The aim of this study was to investigate the growth and lipid accumulation of a new isolated marine microalgal strain by optimizing culture medium composition and applying different stressful culture conditions. Microalga CTM 20023 was isolated from the evaporating salt-ponds at Sfax, Tunisia, using serial-dilution technique from enriched cultures. Phylogenetic analysis based on SSU rDNA and rbcL-3P sequences attributed this isolate to a new species of the Amphora genus. This wild strain possesses rapid gravity sedimentation of 2.91 m h(-1), suitable for an easy and low-cost biomass harvest. The optimization of the composition of the culture medium through statistical experimental designs improved the specific growth rate of Amphora sp. from 0.149 to 0.262 day(-1) and increased its 15-day culture biomass production from 465 to 2200 mg L(-1) (dw) and its lipid content from 140 to 370 mg g(-1) (dw). Highest biomass productivity of 178 mg L(-1) day(-1) was achieved at the 10th day of culture. Highest lipid content of 530 mg g(-1) (dw) was obtained under phosphorus starvation and 64.34% of these lipids were saturated fatty acids. A first growth stage, in optimized condition, would thus offer the maximum productivity for an algal biomass feed stream, followed by second stressful stage for lipid accumulation, thus suitable for biodiesel production.

Optimisation of the critical medium components for better growth of Picochlorum sp. and the role of stressful environments for higher lipid production.

BACKGROUND: Coastal countries that suffer from a scarcity of water, such as Tunisia, have to cultivate marine microalgae on non-arable land in order to produce feedstock and overcome their demands of nutrition and energy. In this framework, a green microalga, CTM 20019, was isolated, identified as Picochlorum sp. and tested for its lipid production. RESULTS: The dry weight of Picochlorum sp. is composed of 163 g kg(-1) lipids, 225 g kg(-1) total sugars, 440 g kg(-1) proteins and 112 g kg(-1) ash rich in potassium, calcium, iron, magnesium and zinc. Gas chromatography-mass spectrometry analysis showed that the main fatty acids were palmitic acid (29%), linolenic acid (26.5%), linoleic acid (23.5%), hexadecatrienoic acid (11%) and hexadecadienoic acid (8.5%). As it is known that culture conditions greatly influence the composition of microalgae, the experiments were designed to optimise the composition of the medium in order to increase Picochlorum sp. growth from OD680nm = 0.53 to OD680nm = 2.2 and lipid accumulation from 163 g kg(-1) to 190 g kg(-1) . The highest lipid contents of 570 and 585 g kg(-1) were achieved under phosphate starvation and sodium carbonate supplementation, respectively. Under these conditions, the fatty acid profile is dominated by mono-unsaturated and polyunsaturated acids, and is therefore suitable for aqua-culture feeding. However, under high salinity, growth and lipid synthesis are inhibited, and the fatty acids are saturate, and the product is therefore suitable for biodiesel. CONCLUSION: This high lipid content rich in essential fatty acids, omega-6 and omega-3, endorses this wild strain of Picochlorum sp. as a promising feedstock for aqua-culture and human nutrition or for the production of biodiesel. © 2013 Society of Chemical Industry.