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This study investigated the effects of gamma ( 137 Cs, 0-250 Gy) and UV (UV-C, 0-12 h) radiation on growth and biodiesel properties of Botryococcus braunii KMITL. For gamma radiation, maximum biomass (1.37 ± 0.02 g L -1 ) was achieved with 50 Gy, while a dose of 200 Gy resulted in the highest hydrocarbon content (51.84 ± 0.20%) and yield (0.66 ± 0.01 g L -1 ). For UV radiation, a 9 h exposure produced the highest biomass (2.45 ± 0.05 g L -1 ), hydrocarbon content (55.01 ± 1.22%), and yield (1.35 ± 0.04 g L -1 ). Algae exposed to gamma radiation within the range of 0-150 Gy exhibited C16:0 as the dominant fatty acid methyl ester (FAME), similar to those exposed to UV radiation, while algae exposed to 200-250 Gy displayed C18:1n9t as the dominant FAME. High levels of gamma and UV radiation were observed to lengthen fatty acid chains and increase unsaturated fatty acids. The cetane values of biodiesel from algae exposed to gamma and UV radiation ranged from 64.55 ± 0.14-66.47 ± 0.20 and 59.43 ± 0.04-65.27 ± 0.22, respectively, all meeting standard criteria. Both gamma and UV radiation also improved the saponification value and cold flow properties of the biodiesel. These findings suggest that controlled levels of gamma and UV radiation effectively enhance hydrocarbon yields with significant implications for biofuel production., (© 2024. The Author(s).)

Rapid light curves are one of the most widely used methods for assessing the physiological state of photosynthetic organisms. While the method has been applied in a range of physiological studies over the last 20 years, little progress has been made in adapting it for the new age of multi-parametric phenotyping. In order to advance research that is aimed at evaluating the physiological impact of multiple factors, the Phenoplate was developed: a simultaneous assessment of temperature and light gradients. It was used to measure rapid light curves of three marine microalgae across a temperature gradient and altered phosphate availability. The results revealed that activation of photoprotective mechanisms occurred with high efficiency at lower temperatures, and relaxation of photoprotection was negatively impacted above a certain temperature threshold in Tetraselmis sp. It was observed that Thalassiosira pseudonana and Nannochloropsis oceanica exhibited two unique delayed non-photochemical quenching signatures: in combinations of low light with low temperature, and darkness with high temperature, respectively. These findings demonstrate that the Phenoplate approach can be used as a rapid and simple tool to gain insight into the photobiology of microalgae., (Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.)

Algal biofuels are a promising alternative to fossil fuels, but their widespread use is hindered by problems with mass production. Light-emitting diodes (LEDs) with specific light wavelengths could be used as an energy source for algal growth and lipid synthesis. In this study, the effects of light source on the biomass and lipid production of the aerial microalga Coccomyxa sp. KGU-D001 were evaluated using LEDs. The integration of two-phase cultures, including growth and lipid production under the stress of nitrate depletion, was assessed for efficient lipid production under liquid- or aerial-phase conditions. Different wavelengths of light (blue, green, and red) were tested under liquid- and aerial-phase conditions. Under aerial-phase culture, the fatty acid contents in biofilm reached 320 mg g DWC -1 with the red LEDs. In view of these findings, we describe a one-step culture method for growth and lipid accumulation in algal biofilm under aerial-phase culture with red LED irradiation. When Coccomyxa biofilm was cultured on wet cotton wool with BBM in a petri dish under the red LED, it was able to grow and accumulate lipids under the aerial-phase condition. Based on the results of this study, a potential method for a continuous biodiesel production system is proposed., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Background: The green microalga Dunaliella salina accumulates a high proportion of β-carotene during abiotic stress conditions. To better understand the intracellular flux distribution leading to carotenoid accumulation, this work aimed at reconstructing a carbon core metabolic network for D. salina CCAP 19/18 based on the recently published nuclear genome and its validation with experimental observations and literature data., Results: The reconstruction resulted in a network model with 221 reactions and 212 metabolites within three compartments: cytosol, chloroplast and mitochondrion. The network was implemented in the MATLAB toolbox CellNetAnalyzer and checked for feasibility. Furthermore, a flux balance analysis was carried out for different light and nutrient uptake rates. The comparison of the experimental knowledge with the model prediction revealed that the results of the stoichiometric network analysis are plausible and in good agreement with the observed behavior. Accordingly, our model provides an excellent tool for investigating the carbon core metabolism of D. salina., Conclusions: The reconstructed metabolic network of D. salina presented in this work is able to predict the biological behavior under light and nutrient stress and will lead to an improved process understanding for the optimized production of high-value products in microalgae.

The unicellular green microalga Haematococcus pluvialis accumulates large amounts of the red ketocarotenoid astaxanthin. Aiming to cultivate these microalgae with high astaxanthin efficiency, cultivations were scaled-up from 1000 mL bottle to 2 L and 8 L airlift photobioreactor using volumetric power consumption rate (W/m 3 ) as scale up strategy. After cultivations, computational fluid dynamics (CFD) simulation was used to investigate the flow patterns, mixing efficiency and gas holdup profile within the 2 L photobioreactor. At the end, astaxanthin content was enhanced with increasing the cultivation volume and highest astaxanthin amount of 49.39 ± 1.64 mg/g cell was obtained in 8 L photobioreactor. Hydrodynamic characteristics of photobioreactor was simulated and gas holdup showed difference between the riser and the downcomer regions. Velocity profiles of air and medium had higher values inside the draft tube than obtained in downcomer region. However liquid circulation was achieved from draft tube to the downcomer, mixing was not provided effectively considering the turbulence kinetic energy. For the further research, some developments about column configuration, sparger diameter may be necessary to enhance the mixing characteristics., (Copyright © 2019 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)

Current research aimed to increase mixotrophic biomass from various organic carbon sources by exploring best light conditions. Three substrates glucose, acetic acid and glycerol were studied for their effects on mixotrophic microalgae cultivation under four light conditions. Light irradiance exhibited variability in growth response and photosynthetic efficiency based on type of substrates used in mixotrophic growth. Each substrate showed variability in light requirements for their effective assimilations. From growth responses, glucose and acetic acid respectively exhibited heterotrophic and mixotrophic (better growth in light) natures. Continuous light-deficient condition was adequate for effective mixotrophic growth as well as energy saving for glucose. However, light-sufficient condition required for effective acetic acid supported mixotrophic growth. Mixotrophic benefits from glycerol and its uptake by Chlorella protothecoides was negligible in all light conditions. Investigation of heterotrophic biomass contribution by various substrates in overall mixotrophic yield, glucose offered maximum approx. 43% contribution., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Melatonin (MLT), a ubiquitously distributed small molecule, functions in plant responses to various biotic and abiotic stresses. However, the interactions between melatonin and other important molecules in Haematococcus pluvialis response stresses are largely unknown. In the present study, exogenous melatonin improved H. pluvialis resistance to nitrogen starvation and high light. We concluded that exogenous melatonin treatment prevented the reactive oxygen species (ROS) burst and limited cell damage induced by abiotic stress through activation of antioxidant enzymes and antioxidants. Astaxanthin, a major antioxidant in H. pluvialis cells, exhibited a 2.25-fold increase in content after treatment with melatonin. The maximal astaxanthin content was 32.4 mg g -1 . The functional roles of the nitric oxide (NO)-mediated mitogen activated protein kinase (MAPK) signaling pathway and cyclic adenosine monophosphate (cAMP) signaling pathway induced by melatonin were also evaluated. The results clearly indicate that cAMP signaling pathways are positively associated with microalgal astaxanthin biosynthesis. Additionally, the NO-dependent MAPK signaling cascade is activated in response to astaxanthin accumulation induced by melatonin, confirming that MAPK is a target of NO action in physiological processes. This work is the first to use H. pluvialis as in vivo model and documents the influence of melatonin on the physiological response to abiotic stress in this microalgae.

In this study, the effect of abiotic stress on the acidophilic eukaryotic microalga, Coccomyxa onubensis, was analyzed for the production of lutein and PUFAs (polyunsaturated fatty acids). It grows autotrophically at a pH of 2.5. It showed a growth rate of 0.30 d -1 , and produced approximately 122.50 mg·L -1 ·d -1 biomass, containing lipids (300.39 mg g -1 dw), lutein (5.30 mg g -1 dw), and β-carotene (1.20 mg g -1 dw). The fatty acid methyl ester (FAME) fraction was 89.70 mg g -1 dw with abundant palmitic acid (28.70%) and linoleic acid (37.80%). The addition of 100 mM NaCl improved the growth rate (0.54 d -1 ), biomass productivity (243.75 mg·L -1 ·d -1 ), and lipids accumulation (416.16 mg g -1 dw). The microalga showed a lutein content of 6.70 mg g -1 dw and FAME fraction of 118.90 mg g -1 dw; 68% of the FAMEs were PUFAs. However, when 200-500 mM salt was added, its growth was inhibited but there was a significant induction of lutein (up to 7.80 mg g -1 dw). Under continuous illumination with PAR (photosynthetically active radiations) +UVA (ultraviolet A, 8.7 W m -2 ), C. onubensis showed a growth rate of 0.40 d -1 , and produced 226.3 mg·L -1 ·d -1 biomass, containing lipids, (487.26 mg g -1 dw), lutein (7.07 mg g -1 dw), and FAMEs (232.9 mg g -1 dw); 48.4% of the FAME were PUFAs. The illumination with PAR + UVB (ultraviolet B, 0.16 W m -2 ) was toxic for cells. These results indicate that C. onubensis biomass is suitable as a supplement for functional foods and/or source of high added value products., (Copyright © 2018 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)

The study of microalgal culture has been growing in recent decades, because the cellular structure of microalgae has diverse highly valuable metabolites that have attract attention of numerous companies and research groups. The pigment astaxanthin is considered one of the most powerful antioxidants in nature. The microalga Haematococcus pluvialis was proposed as one of the best natural astaxanthin sources, because it can accumulate high amount of the pigment. In this work, we studied different stress treatments on H. pluvialis growth cultures as well as astaxanthin production under autotrophic growth conditions. The results showed that extending nitrogen starvation before increasing radiation intensity up to 110 μmol photons m -2 s -1 during late the palmella cell phase incremented the astaxanthin concentration up to 2.7% of dry biomass with an efficient light energy utilization during the stress stage.

Aerial microalgae are more exposed to harsh and rapidly changing environmental conditions, including desiccation and radiation. Under high light, aerial algae in the desiccated state would be highly subject to photodamage. Therefore, aerial algae need effective protective mechanisms to dissipate excess excitation energy. In this study, the changes in photosynthetic behaviors during desiccation and after rehydration in Trentepohlia jolithus were confirmed using chlorophyll a fluorescence (OJIP) transient, allowing determination of the photoprotection mechanisms of this aerial alga. The filaments of T. jolithus cells at 25% relative air humidity (RH) are significantly shrunken compared with those at 100% and 87% RH, decreasing the surface area for light absorption. At 25% RH, the shape and intensity of the OJIP transient disappeared, but recovered rapidly to the level at 100% RH after 5 s of rehydration. Compared with 100% RH, the maximum quantum yield of PSII (φ Po ), phenomenological energy fluxes for absorption (ABS/CSm) and active PSII reaction centers (RCs) at 25% RH decreased significantly, the specific energy fluxes for absorption (ABS/RC) increased significantly, but the specific energy fluxes for trapping (TRo/RC) at 25% RH did not change. These parameters at 25% RH recovered rapidly to the level at 100% RH after 5 s of rehydration. These results suggest that the efficiency of PSII light absorption and activities of PSII RCs were reversibly down-regulated in desiccated T. jolithus, which may be a special adaptive mechanism for the survivability of aerial microalgae in habitats with rapidly changing water availability., (© 2016 German Botanical Society and The Royal Botanical Society of the Netherlands.)

The effects of light-emitting diode (LED) wavelength, light intensity, nitrate concentration, and time of exposure to different LED wavelength stresses in a two-phase culture on lipid production were evaluated in the microalga, Picochlorum atomus. The biomass produced by red LED light was higher than that produced by purple, blue, green, or yellow LED and fluorescent lights from first phase of two-phase culture. The highest lipid production of P. atomus was 50.3% (w/w) with green LED light at 2days of second phase as light stress. Fatty acid analysis of the microalgae showed that palmitic acid (C16:0) and linolenic acid (C18:3) accounted for 84-88% (w/w) of total fatty acids from P. atomus. The two-phase culture of P. atomus is suitable for biofuel production due to higher lipid productivity and favorable fatty acid composition., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

In this study, an alternative approach was proposed where excess seawater would be added only during inoculation (DD) rather than daily addition (FD). Growth and metabolite contents of Tetraselmis sp. weren't affected for daily increase of 2% NaCl salinity. Tetraselmis sp. was then cultured in DD and FD pond. In DD pond, initial culture depth was 23.5cm and its depth reduced as no water was added; for FD pond, everyday sterilized seawater was added to maintain 20cm depth. DD pond had higher biomass productivity compared to FD pond, until DD pond was deeper than FD pond; metabolite content and FAME profile of Tetraselmis sp. were also similar in both cultures. Therefore, considering the simplicity in operation, halo tolerant microalgae can be grown in DD pond method., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Unicellular green microalgae Haematococcus pluvialis was mutated with (60)Co-γ irradiation to promote growth rate and increase astaxanthin yield under high concentration of CO2 stress. The average specific growth rate of H. pluvialis mutated with 4000 Gy γ-ray irradiation was increased by 15% compared with the original strain with air aeration. The mutant grew best with 6% CO2 (the maximum specific growth rate was 0.60/d) when it was cultured with high concentrations of CO2 (2-10%). The peak biomass productivity (0.16 g/L/d) of the mutant cultured with 6% CO2 was 82% higher than that of the mutant with air. The astaxanthin yield and lipid content of the mutant induced with 6% CO2 and high light (108 μmol photons m(-2) s(-1)) increased to 46.0mg/L and 45.9%, which were 2.4 and 1.3 times higher than those of the wild-type strain, respectively., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

It has been reported in the literature that exposure of microalgae cells to audible sound could promote growth. This study examined the effect of sound waves with the frequency of 1100 Hz, 2200 Hz, and 3300 Hz to stimulate the biomass productivity of an Oklahoma native strain, Picochlorum oklahomensis (PO). The effect of the frequency of sound on biomass mass was measured. This study demonstrated that audible sound treatment of the algae cultures at 2200 Hz was the most effective in terms of biomass production and volumetric oil yield., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Carotenoids prevent different degenerative diseases and improve human health. Microalgae are commercially exploited for carotenoids, including astaxanthin and β-carotene. Two commercially important microalgae, Dunaliella salina and Tetraselmis suecica, were treated with plant hormones salicylic acid (SA) and methyl jasmonate (MJ), or by UV-C radiation (T. suecica only) and a combination thereof. Significant increases in total carotenoids were found for D. salina and T. suecica after treatment with MJ (10 μmol/L) and SA (70-250 μmol/L), respectively. T. suecica also had significant increases in total carotenoids following UV-C radiation compared to control cultures. Among the carotenoids, lutein was the highest induced carotenoid. A combination of these two treatments also showed a significant increase in total carotenoids and lutein for T. suecica, when compared to controls. Plant hormones and UV-C radiation may be useful tools for increasing carotenoid accumulation in green microalgae although the responses are species- and dose-specific and should be trialed in medium to large scale to explore commercial production.

The marine microalga Tetraselmis subcordiformis could photoproduce hydrogen under the regulation of carbonyl cyanide m-chlorophenylhydrazone (CCCP), and a hydrogen production process kinetic analysis was characterized by two peaks, suggesting that two distinct mechanisms might exist in this alga. Therefore, 2D nanoliquid chromatography-tandem mass spectrometry (LC-MS/MS) was introduced to analyze the proteome of samples from different time points. A total of 912 proteins were identified, providing a global view of the cellular responses at the proteomic level. These proteins can be divided into multiple functional groups including stress responses, energy metabolism and redox homeostasis. The quantitative proteomic data provided more details on the electron donors for hydrogen production. During the first stage, photosystem II produced electrons for hydrogen production; during the second stage, metabolites were the major electron donors via nonphotochemical plastoquinone reduction by NADH dehydrogenase.

We recently evaluated the relationship between abiotic environmental stresses and lutein biosynthesis in the green microalga Dunaliella salina and suggested a rational design of stress-driven adaptive evolution experiments for carotenoids production in microalgae. Here, we summarize our recent findings regarding the biotechnological production of carotenoids from microalgae and outline emerging technology in this field. Carotenoid metabolic pathways are characterized in several representative algal species as they pave the way for biotechnology development. The adaptive evolution strategy is highlighted in connection with enhanced growth rate and carotenoid metabolism. In addition, available genetic modification tools are described, with emphasis on model species. A brief discussion on the role of lights as limiting factors in carotenoid production in microalgae is also included. Overall, our analysis suggests that light-driven metabolism and the photosynthetic efficiency of microalgae in photobioreactors are the main bottlenecks in enhancing biotechnological potential of carotenoid production from microalgae.

The optimal conditions for cultivating the thermotolerant lipid-rich microalga Desmodesmus sp. F2 to achieve maximal lipid productivity were determined in this study. The conditions were light intensity, 700μmol/m(2)s; temperature, 35°C; cultivation nitrogen source, nitrate; initial nitrogen level, 6.6mM nitrogen. Carbon dioxide (2.5%, 0.2 vvm) was pumped into the cultures continuously. In the pre-optimized conditions, the maximal lipid productivity of this microalga was 113mg/L/d, which was raised to 263mg/L/d in the optimized conditions. This level of lipid productivity of microalgae is the highest ever reported in the literature. Fatty acid composition of the lipid produced by Desmodesmus sp. F2 in the optimal conditions was determined, in which C16 and C18 species accounted for 95% of the fatty acids. Saturated, monounsaturated and polyunsaturated fatty acids accounted for 38.9%, 33.1% and 22.6%, respectively. Based on the analysis, this lipid quality makes it a good feedstock for biodiesel production., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Specific growth rates and hydrocarbon contents of Botryococcus braunii strain Showa were measured under a wide range of CO2, salinity, temperature, and irradiance conditions. The bubbling CO2 concentration of 0.2-5% and no addition of salinity were favorable conditions for growth. The strain cannot grow at 5°C and above 35°C under any irradiance levels. Maximum specific growth rate of 0.5 day(-1) (doubling time of 1.4 days), the highest value reported for B. braunii in the past studies, was observed at 30°C and 850 μmol photons m(-2) s(-1). Since hydrocarbon productivity, shown as the product of hydrocarbon content and specific growth rate, increased with the increasing specific growth rate, we conclude that more efficient hydrocarbon production by the mass culture of strain Showa can be achieved by maintaining higher specific growth rate based on the culture conditions presented in this study., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

The aim of this study was to test three flat-plate photobioreactor configurations for cultivation of marine green alga Dunaliella tertiolecta under non-axenic growth conditions and to characterize and quantify the associated bacteria. The photobioreactor cultivations were conducted using tap water-based media. Static mixers intended to enhance mixing and light utilization did not generally increase algal growth at the low light intensities used. The maximum biomass concentration (measured as volatile suspended solids) and maximum specific growth rate achieved in the flat plate with no mixer were 2.9 g l⁻¹ and 1.3 day⁻¹, respectively. Based on quantitative polymerase chain reaction, bacterial growth followed the growth of D. tertiolecta. Based on 16S rDNA amplification and denaturing gradient gel electrophoresis profiling, heterotrophic bacteria in the D. tertiolecta cultures mainly originated from the non-axenic algal inocula, and tap water heterotrophs were not enriched in high chloride media (3 % salinity). Bacterial communities were relatively stable and reproducible in all flat-plate cultivations and were dominated by Gammaproteobacteria, Flavobacteria, and Alphaproteobacteria.

Determination of organic acids in intracellular extracts and in the cultivation media of marine microalgae aid investigations about metabolic routes related to assimilation of atmospheric carbon by these organisms, which are known by their role in the carbon dioxide sink. The separation of these acids was investigated by hydrophilic interaction liquid chromatography (HILIC) using isocratic elution with a mobile phase composed of 70:30 v/v acetonitrile/20 mmol/L ammonium acetate buffer (pH 6.8) and detection at 220 nm. HILIC allowed the determinations of glycolic acid, the most important metabolite for the evaluation of the photorespiration process in algae, to be made with better selectivity than that achieved by reversed phase liquid chromatography, but with less detectability. The concentration of glycolic acid was determined in the cultivation media and in intracellular extracts of the algae Tetraselmis gracilis and Phaeodactylum tricornutum submitted to different conditions of aeration: (i) without forced aeration, (ii) aeration with atmospheric air, and (iii) bubbling with N(2). The concentration of glycolic acid had a higher increase as the cultures were aerated with nitrogen, showing higher photorespiratory flux than that occurring in the cultures aerated with atmospheric air., (Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)