• Journal of the Korea Academia-Industrial cooperation Society
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• v.10 no.1
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• pp.194-199
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• 2009

An efficient production method of food-grade heamatococcus extract was developed based on stepwise enzymatic hydrolysis. In the first step, Haematococcus pluvialis cells hydrolysis carried out with commercially available exopeptidase(Flavourzyme) and endopeptidase (Alcalase), resulted in increased astaxanthin content. In the second step, proteolytic hydrolyzed H. pluvialis cells treated with hetero-polysaccharides hydrolytic enzyme (Viscozyme). By two-stage treatments using Alcalase and Flavourzyme and Viscozyme, the highest astaxanthin content was obtained. The astaxanthin content was remarkably enhanced by 320% $(529{\mu}g/g\rightarrow2,256{\mu}g/g)$ than that of the non-treated extract. And then, antioxidative activities determined by DPPH method were increased with increasing the astaxanthin content in haematococcus extract prepared by enzymatic hydrolysis.

• ALGAE
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• v.38 no.1
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• pp.1-22
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• 2023

An enormous body of research is focused on finding ways to commercialize carotenoids produced by the unicellular green alga, Haematococcus, often without the benefit of a sound phylogenetic assessment. Evidence of cryptic diversity in the genus means that comparing results of pigment studies may be confounded by the absence of a phylogenetic framework. Moreover, previous work has identified unnamed strains that are likely candidates for species status. We reconstructed the phylogeny of an expanded sampling of Haematococcus isolates utilizing data from nuclear ribosomal markers (18S rRNA gene, 26S rRNA gene, internal transcribed spacer [ITS]-1, 5.8S rRNA gene, and ITS-2) and the rbcL gene. In addition, we gathered morphological, ultrastructural and pigment data from key isolates of Haematococcus. Our expanded data and taxon sampling support the concept of a new species, H. privus, found exclusively in North America. Despite overlap in numerous morphological traits, results indicate that ratios of protoplast length to width and akinete diameter may be useful for discriminating Haematococcus lineages. High growth rate and robust astaxanthin yield indicate that H. rubicundus (SAG 34-1c) is worthy of additional scrutiny as a pigment source. With the description of H. privus, the evidence supports the existence of at least five, species-level lineages in the genus. Our phylogenetic assessment provides the tools to frame future pigment investigations of Haematococcus in an updated evolutionary context. In addition, our investigation highlighted open questions regarding polyploidy and sexuality in Haematococcus which demonstrate that much remains to be discovered about this green flagellate.

• Journal of Microbiology and Biotechnology
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• v.16 no.6
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• pp.821-831
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• 2006

Unicellular green algae of the genus Haematococcus have been studied extensively as model organisms for secondary carotenoid accumulation. Upon environmental stress, such as strong irradiance or nitrogen deficiency, unicellular green algae of the genus Haematococcus accumulate secondary carotenoids in vesicles in the cytosol. Because secondary carotenoid accumulation occurs only upon specific environmental stimuli, there is speculation about the regulation of the biosynthetic pathway specific for secondary carotenogenesis. Because the carotenoid biosynthesis pathway is located both in the chloroplast and the cytosol, communication between both cellular compartments must be considered. Recently, the induction and regulation of astaxanthin biosynthesis in microalgae received considerable attention because of the increasing use of this secondary carotenoid as a source of pigmentation for fish aquaculture, as a component in cancer prevention, and as a free-radical quencher. This review summarizes the biosynthesis and regulation of the pathway, as well as the biotechnology of astaxanthin production in Haematococcus.

• Microbiology and Biotechnology Letters
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• v.48 no.3
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• pp.337-343
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• 2020

In this study, the medium optimization for cell growth and metabolite formation of Haematococcus sp. under mixotrophic cultivation was investigated. As a result, modified MS medium was selected as the basal medium; glucose was selected as the carbon source, with an optimum concentration of 10 g/l, and potassium nitrate was chosen as the nitrogen source, with an optimum concentration of 1.9 g/l. Under optimum conditions, Haematococcus sp. demonstrated an increase in biomass concentration from 0.18 gDW/l to 5.58 gDW/l in 14 days, after which there was a 31-fold increase in its growth. At the same time, the concentrations of chlorophyll and carotenoids were 172.16 mg/l and 42.33 mg/l, respectively. This work will contribute to the basic data for mass cultivation of microalgae.

• KSBB Journal
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• v.26 no.5
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• pp.381-385
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• 2011

Studies were made to improve the stability of astaxanthin which has application limitations caused by light and thermal stability problems in spite of its strong anti-oxidant property. Astaxanthin was extracted from Haematococcus pluvialis with supercritical carbon dioxide. Liposomal encapsulation of astaxanthin to improve the stability was made with high pressure homogenizer. The narrow size distribution was observed with astaxanthin liposomes. Tests on light and thermal stabilities resulted that the liposormal encapsulation improved the stability of astaxanthin for cosmeceutical purposes.

• Journal of Microbiology and Biotechnology
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• v.25 no.2
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• pp.238-246
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• 2015

A unicellular red microalga was isolated from environmental freshwater in Korea, and its morphological, molecular, and biochemical properties were characterized. Morphological analysis revealed that the isolate was a unicellular biflagellated green microalga that formed a non-motile, thick-walled palmelloid or red aplanospore. To determine the taxonomical position of the isolate, its 18S rRNA and rbcL genes were sequenced and phylogenetic analysis was performed. We found that the isolate was clustered together with other related Haematococcus strains showing differences in the rbcL gene. Therefore, the isolated microalga was classified into the genus Haematococcus, and finally designated Haematococcus sp. KORDI03. The microalga could be cultivated in various culture media under a broad range of pH and temperature conditions. Compositions of the microalgal cellular components were analyzed, and its protein, carbohydrate, and lipid compositions were estimated to be 21.1 ± 0.2%, 48.8 ± 1.8%, and 22.2 ± 0.9%, respectively. In addition, D-glucose and D-mannose were the dominant monosaccharides in the isolate, and its amino acids were composed mainly of aspartic acid, glutamic acid, alanine, and leucine. Moreover, several polyunsaturated fatty acids accounted for about 80% of the total fatty acids in Haematococcus sp. KORDI03, and the astaxanthin content in the red aplanospores was estimated to be 1.8% of the dry cell weight. To the best of our knowledge, this is the first report of an Haematococcus sp. isolated from Korea, which may be used for bioresource production in the microalgal industry.

• Journal of Microbiology and Biotechnology
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• v.16 no.6
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• pp.849-854
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• 2006

The effects of Haematococcus pluvialis extracellular products on microbial growth and bacteriocin production were investigated to improve bacteriocin synthesis during the growth cycle of Lactobacilli. Lactobacillus pentosus KJ-108, L. plantarum KJ-10311, and L. sakei KJ-2008 were cultured in MRS and enriched medium (ERM) with or without supplement of the extracellular products obtained from a late exponential phase culture of Haematococcus pluvialis in modified Bold's basal medium (MBBM). In both MRS and ERM, the extracellular products strongly enhanced the growth as well as the bacteriocin production of all the lactic acid bacteria tested. The enhancing effect was observed in ERM with pH adjusted at 5 and 6. In addition, some difference in growth effects with the extracellular products of H. pluvialis was observed between pH 5 and 6 in ERM, but no effect was observed in the minimal medium. The final biomass and the final concentration of bacteriocin activity were associated with the cell growth that was promoted by the extracellular products of H. pluvialis, and the enhanced cell growth of the three lactic acid bacterial strains induced the increase of the specific bacteriocin production. Therefore, bacteriocin production and activity were influenced by the addition of the extracellular products of H. pluvialis in the culture medium.

• 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.

• ALGAE
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• v.28 no.2
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• pp.185-192
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• 2013

A new cold-adapted Arctic strain of Haematococcus pluvialis from Blomstrandhalv${\o}$ya Island (Svalbard) is described. This strain is predominantly always in non-motile palmelloid stage. Transmission electron microscopy showed the presence of very thick cell wall and abundant lipid vesicles in the palmelloids, including red and green cells. The external morphology of the non-motile palmelloid and motile bi-flagellated cells of our strain is similar to H. pluvialis; however it differs from H. pluvialis in physiology. Our strain is adapted to live and produce astaxanthin in the low temperature ($4-10^{\circ}C$), whilst the usual growth temperature for H. pluvialis is between $20-27^{\circ}C$. Phylogenetic analysis based on 18S rRNA gene data showed that our strain nested within the Haematococcus group, forming a sister relationship to H. lacustris and H. pluvialis, which are considered synonymous. Therefore, we identified our Arctic strain as H. pluvialis.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.181-184
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• 2001

The biological fixation of carbon dioxide using microalgae have many advantages over chemicals and remove carbon dioxide simultaneously. A ketocarotenoid astaxanthin is hyper-accumulated in the green freshwater microalga, Haematococcus pluvialis. In the present study, the combine effects of carbon dioxide concentration and light intensity on the growth of H. pluvilais were investigated. The carbon dioxide concentration above 10% caused a severe inhibition and around 5% is optimal for growth. Adaptation to high concentration of carbon dioxide enhanced the $CO_2$ tolerance. Specific growth rate calculated differently based upon cell number or dry weight because of the distinctive life cycle patterns of H. pluvialis : small-sized motile green cell and thick cell walled red cyst cell. Based on the light dependence of H. pluvialis, internally illuminated air-lift photobioreactor was designed and operated. Gradual increase of light supply gave more active growth and more effective productivity of astaxanthin than constant light supply.