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Carotenoids are hydrophobic pigments produced exclusively by plants, fungi, and specific microbes. Microalgae are well suited for the production of valuable carotenoids due to their rapid growth, efficient isoprenoid production pathway, and ability to store these compounds within their cells. The possible markets for bio-products range from feed additives in aquaculture and agriculture to pharmaceutical uses. The production of carotenoids in microalgae is affected by several environmental conditions, which can be utilized to enhance productivity. The current study focused on optimizing the extraction parameters (time, temperature, and extraction number) to maximize the yield of carotenoids. Additionally, the impact of various nitrogen sources (ammonia, nitrate, nitrite, and urea) on the production of lutein and loroxanthin in Scenedesmus obliquus was examined. To isolate the carotenoids, 0.20 g of biomass was added to 0.20 g of CaCO3 and 10.0 mL of ethanol solution containing 0.01% (w/v) pyrogallol. Subsequently, the extraction was performed using an ultrasonic bath for a duration of 10 min at a temperature of 30 °C. This was followed by a four-hour saponification process using a 10% methanolic KOH solution. The concentration of lutein and loroxanthin was measured using HPLC–DAD at 446 nm, with a flow rate of 1.0 mL/min using a Waters YMC C30 Carotenoid column (4.6 × 250 mm, 5 μm). The confirmation of carotenoids after their isolation using preparative chromatography was achieved using liquid chromatography–tandem mass spectrometry (LC–MS/MS) with an atmospheric pressure chemical ionization (APCI) probe and UV–vis spectroscopy. In summary, S. obliquus shows significant promise for the large-scale extraction of lutein and loroxanthin. The findings of this study provide strong support for the application of this technology to other species. [ABSTRACT FROM AUTHOR]

Microalgae produce a variety of high-value chemicals including carotenoids. Fucoxanthin is also a carotenoid that has many physiological functions and biological properties. For this reason, the cost-effective production of fucoxanthin at an industrial scale has gained significant attention. In the proposed study, fucoxanthin production was aimed to be increased by altering the culture conditions of N. shiloi. The effect of light intensity aeration rate, different nitrogen sources, and oxidative stress on the biomass and fucoxanthin productivity have been discussed. Based on these results, the fucoxanthin increased to 97.45 ± 2.64 mg/g by adjusting the light intensity to 50 µmol/m2s, and aeration rate at 5 L/min using oxidative stress through the addition of 0.1 mM H2O2 and 0.1 mM NaOCl to the culture medium. Fucoxanthin was then purified with preparative HPLC using C30 carotenoid column (10 mm × 250 mm, 5 μm). After the purification procedure, Liquid chromatography tandem mass spectrometry (LC–MS/MS) and UV-vis spectroscopy were employed for the confirmation of fucoxanthin. This study presented a protocol for obtaining and purifying considerable amounts of biomass and fucoxanthin from diatom by manipulating culture conditions. With the developed methodology, N. shiloi could be evaluated as a promising source of fucoxanthin at the industrial scale for food, feed, cosmetic, and pharmaceutical industries. [ABSTRACT FROM AUTHOR]

Microalgae with their improved growth rates and accumulation of high-value-added products make their commercial production attractive. Among them, lutein, which is a carotenoid, plays a very important role due to its various applications in the food and pharmaceutical industry. Induction of its biosynthesis can be triggered by various stress conditions like light. In this study, three different light intensities (50,150 and 300 µmol photons/m²s) and aeration rates (1, 3, and 5 L/min) were utilized to induce the lutein biosynthesis and biomass productivity in Scenedesmus obliquus. Lutein was isolated by preparative chromatography using a semiprep C30 column (10 × 250 mm, 5µm) and its confirmation was made by LC-MS/MS. According to the results, Scenedesmus obliquus synthesized the maximum lutein (8.01 ± 0.1 mg/g) with biomass productivity of 1.698 g/L at 150 µmol photons/m²s light intensity using 3 L/min as aeration rate. To the best of the authors' knowledge, this was the first study that the lutein was isolated by preparative chromatography using semiprep C30 carotenoid column with a simple and rapid separation, which can be used as a reference methodology for the isolation of other carotenoids. Scenedesmus obliquus can be an important alternative source for commercial production of lutein, as it is indicated from the results of this study. [ABSTRACT FROM AUTHOR]

Microalgae with high growth rates represent a potential source of carotenoids including fucoxanthin. For commercial fucoxanthin production, choosing the cultivation system is a critical parameter where the final biomass must contain a high content of the target product. To realize this, the reaction system must allow the use of stress conditions that induce the synthesis of these compounds. In this study, oxidative stress was applied to the diatom Amphora capitellata by the formation of reactive oxygen species in the culture medium through chemical reactions which generate. The effect of this oxidative stress on the amount of fucoxanthin and specific growth rate of A. capitellata was investigated. In the presence of H2O2/NaOCl, fucoxanthin content was 41.83 ± 0.92 mg g−1 as determined by HPLC–DAD a 2.20-fold increase compared with the control group. For the purification of fucoxanthin, preparative HPLC using a semi-prep C30 carotenoid column (10 × 250 mm, 5 µm) was used for the first time with an isocratic elution of 75:25 (methanol:acetonitrile) mobile phase at a flow rate of 4 mL min−1. Liquid chromatography tandem mass spectrometry (LC–MS/MS) was employed for the confirmation of fucoxanthin. The developed method has potential to obtain highly pure fucoxanthin for food and pharmaceutical applications using A. capitellata as a commercial source of fucoxanthin, particularly after its enhancement via oxidative stress. [ABSTRACT FROM AUTHOR]