• Journal of Marine Bioscience and Biotechnology
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• v.6 no.1
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• pp.31-40
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• 2014

The production of astaxanthin in the microalga Haematococcus pluvialis has been investigated using a sequential methodology based on the application of two types of statistical designs. The employed preliminary experiment was a fractional factorial design $2^6$ in which the factors studied were: excessive irradiance and nitrate starvation, phosphate deficiency, acetate supplementation, salt stress, and elevated temperature. The experimental results indicate that the amount of astaxanthin accumulation in the cells can be enhanced by excessive irradiance and nitrate starvation whereas the other factors tested did not yield any enhancement. In the subsequent experiment, a central composite design was applied with four variables, light intensity, nitrate, phosphate, and acetate, at five levels each. The optimal conditions for the highest astaxanthin production were found to be $1040{\mu}E/(m^2{\cdot}s)$ light intensity, 0.04 g/L nitrate, 0.31 g/L phosphate, 0.05 g/L acetate concentration.

• Korean Chemical Engineering Research
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• v.44 no.1
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• pp.46-51
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• 2006

As one of the $CO_2$ reduction strategies, a biological method was proposed to convert $CO_2$ to useful biomass with antioxidant carotenoids by photosynthetic microorganisms. One of the photoautotrophs, Haematococcus pluvialis is a freshwater green microalga and accumulates the secondary carotenoid astaxanthin during induction of green vegetative cells to red cyst cells. In this study, $CO_2$ fixation and astaxanthin production using H. pluvialis was conducted by photoautotrophic culture in the $CO_2$ supplemented photo-incubator. Maximum growth rate of H. pluvialis was obtained at a 5％ $CO_2$ environment on basic N and P conditions of NIES-C medium. The photoautotrophic induction consisted of 5％ $CO_2$ supply and high light illumination promoted astaxanthin synthesis in H. pluvialis, yielding an astaxanthin productivity of $9.6mg/L{\cdot}day$ and a $CO_2$ conversion rate of $27.8mg/L{\cdot}day$ to astaxanthin. From the results the sequential photoautotrophic culture and induction process using H. pluvialis is expecting an alternative $CO_2$ reduction technology with a function of valuable biosubstance production.

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.10
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• pp.1363-1368
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• 2008

The extraction and quantitative analysis conditions for astaxanthin from Haematococcus pluvialis, and the structural characteristics of H. pluvialis extract, H. pluvialis hydrolysate and synthetic astaxanthin were investigated using UV/visible and FT-IR spectrometers. Astaxanthin was dissolved in methanol, and then treated to enhance the solubility by sonication for 45 min. With sonication pretreatment, the solubility of astaxanthin increased up to 1.5 times compared to that without sonication. The extracts were hydrolyzed by cholesterol esterase for the analysis of H. pluvialis extract containing astaxanthin ester. A HPLC method using reverse phase C18 column with methanol-water (95:5, v/v) as mobile phase was developed to analyze astaxanthin. After hydrolysis, the absorption spectrum of H. pluvialis hydrolysat was changed to similar pattern to synthetic astaxanthin, confirming the extraction and analysis condition of astaxanthin from H. pluvialis.

• Applied Biological Chemistry
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• v.51 no.3
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• pp.247-249
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• 2008

• Journal of the Korea Academia-Industrial cooperation Society
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• v.21 no.10
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• pp.64-71
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• 2020

This study evaluated the effects of the culture media and light sources on the growth of microalgae Haematococuus pluvialis. Limited ingredient medium, Modified Bold's Basic Medium (MBBM), commercial liquid fertilizer medium Neo, and seven different light sources with different wavelengths were used to incubate H. pluvialis for 39 days, and the growth rates were compared. As a result, the growth of H. pluvialis, a limited ingredient medium, produced the highest cell growth in the fluorescent light source while cell growth was the lowest in the blue+red LED. The growth of H. pluvialis in commercial medium Neo was highest in the fluorescent light source, and cell growth was lowest in the blue LED. In this study, the MBBM culture medium showed better results than the Neo culture medium. Microalgae grown in the fluorescent light source using the MBBM culture medium showed the best cell growth result in this study. The results were optimized for the culture medium, light source, and light quantity in H. pluvialis culture for the production of secondary metabolites and provide basic data for the mass culture of microalgae.

• Microbiology and Biotechnology Letters
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• v.35 no.4
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• pp.292-297
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• 2007

The unicellular green alga, Haematococcus pluvialis used as a biological production system for astaxanthin. It accumulates large amounts of the red ketocarotenoid astaxanthin when exposed to various environmental stress such as active oxygen species and high light intensities. To induce astaxanthin biosynthesis of H. pluvialis, cells were incubated in either nitrate free at $25^{\circ}C$ under continuous high light intensity ($1,000\;{\mu}mol$ photons $m^{-2}s^{-1}$) for 2 days or high light stress only. Expressions of astaxanthin biosynthetic genes such as carotenoid hydroxylase, IPP isomerase and ${\beta}$-carotene ketolase were monitored under different culture conditions by using real time RT-PCR. All the subjected genes increased their expression under highlight and N-deprivation condition where a large amount of astaxanthin was accumulated.

• ALGAE
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• v.28 no.2
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• pp.193-202
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• 2013

Nitrogen availability and cell density each affects growth and cellular astaxanthin content of Haematococcus pluvialis, but possible combined effects of these two factors on the content and productivity of astaxanthin, especially under outdoor culture conditions, is less understood. In this study, the effects of the initial biomass densities IBDs of 0.1, 0.5, 0.8, 1.5, 2.7, 3.5, and 5.0 g $L^{-1}$ DW and initial nitrogen concentrations of 0, 4.4, 8.8, and 17.6 mM nitrate on growth and cellular astaxanthin content of H. pluvialis Flotow K-0084 were investigated in outdoor glass column photobioreactors in a batch culture mode. A low IBD of 0.1 g $L^{-1}$ DW led to photo-bleaching of the culture within 1-2 days. When the IBD was 0.5 g $L^{-1}$ and above, the rate at which the increase in biomass density and the astaxanthin content on a per cell basis was higher at lower IBD. When the IBD was optimal (i.e., 0.8 g $L^{-1}$), the maximum astaxanthin content of 3.8% of DW was obtained in the absence of nitrogen, whereas the maximum astaxanthin productivity of 16.0 mg $L^{-1}\;d^{-1}$ was obtained in the same IBD culture containing 4.4 mM nitrogen. The strategies for achieving maximum Haematococcus biomass productivity and for maximum cellular astaxanthin content are discussed.

• ALGAE
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• v.28 no.2
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• pp.131-147
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• 2013

Major progress has been made in the past decade towards understanding of the biosynthesis of red carotenoid astaxanthin and its roles in stress response while exploiting microalgae-based astaxanthin as a potent antioxidant for human health and as a coloring agent for aquaculture applications. In this review, astaxanthin-producing green microalgae are briefly summarized with Haematococcus pluvialis and Chlorella zofingiensis recognized to be the most popular astaxanthin-producers. Two distinct pathways for astaxanthin synthesis along with associated cellular, physiological, and biochemical changes are elucidated using H. pluvialis and C. zofingiensis as the model systems. Interactions between astaxanthin biosynthesis and photosynthesis, fatty acid biosynthesis and enzymatic defense systems are described in the context of multiple lines of defense mechanisms working in concert against photooxidative stress. Major pros and cons of mass cultivation of H. pluvialis and C. zofingiensis in phototrophic, heterotrophic, and mixotrophic culture modes are analyzed. Recent progress in genetic engineering of plants and microalgae for astaxanthin production is presented. Future advancement in microalgal astaxanthin research will depend largely on genome sequencing of H. pluvialis and C. zofingiensis and genetic toolbox development. Continuous effort along the heterotrophic-phototrophic culture mode could lead to major expansion of the microalgal astaxanthin industry.

• Journal of Microbiology and Biotechnology
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• v.11 no.6
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• pp.1024-1030
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• 2001

The key factors for high-density Haematococcus pluvialis cultures and conditions for astaxanthin induction were examined to maximize astaxanthin production. Light intensity was found to be the most important factor, and thus experiments were found to be the most important factor, and thus experiments were carried out using different light sources and intensities. A high cell density of over 2.7 g/l was obtained at $75{\mu}E/m^2/s$, whereas a much lower cell concentration (<1.0 g/ 1) was obtained with lower light intensities $(15-30{\mu}E/m^2/s$. A high light intensity and the supplement of 470 nm photons had a more dramatic effect on the final astaxanthin concentration and per cell astaxanthin content. A maximum astaxanthin concentration of 6.5 mg/l was obtained at a light intensity of $160{\mu}E/m^2/s$, whereas only 1.3 and 0.7 mg/l were obtained at 30 and $15{\mu}E/m^2/s$, respectively. A supplement of 470 nm photons enhanced the carotenoid and chlorophyll formation.

• Journal of Microbiology and Biotechnology
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• v.19 no.11
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• pp.1333-1341
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• 2009

Haematococcus pluvial is, a green alga, accumulates astaxanthin (3,3'-dihydroxy-$\beta$,$\beta$'-carotene-4,4'-dione) upto 2-3% on a dry weight basis. In the present study, identification of carotenoids from Haematococcus cyst cell extract by HPLC and LC-MS (APCI) and their antioxidant properties were evaluated in in vitro model systems. The extract exhibited 89% and 78% antioxidant activities in the $\beta$-carotene linoleate model and the hydroxyl radical scavenging model, at 9 ppm of total carotenoid, respectively. The extract also showed 80%, 85%, and 79% antioxidant activities against lipid peroxidation in the kidney, brain, and liver of rats. Low-density lipoprotein oxidation induced by $Cu^{2+}$ ions was also protected (45%, 64%, and 75%) by the extract in a dose-dependent manner with different carotenoid levels. Thiobarbituric acid reactive substances concentration in the blood, liver, and kidney of rats were also significantly (p<0.005) decreased in H. pluvialis-treated rats. The potent antioxidant activity is attributable to various carotenoids present in the extract.