• ALGAE
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• 제24권2호
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• pp.121-127
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• 2009

The unicellular green alga, Haematococcus pluvialis, accumulates the highest level of astaxanthin among knownastaxanthi.n-producing organisms. Light is the most important factor to induce astaxanthin by H. pluvialis. BIue andred LEDs, whose ${\lambda}_{max}$'s are 470 and 665 nm, respectively, were used for internally illuminated light sources.Fluorescent lamps were also used for both internal and external illumination sources. The astaxanthin levels in thesevarious lighting systems were analyzed and compared each other. The cultures under internally illuminated LEDsaccumulaled 20% more astaxanthin than those under fluorescent lamp. Furthermore, LEDs generated much lessheat than the fluorescent lamps, which gives one more reason for the LEDs being a suitable internal Light source forastaxanthin induction. The results reported here would lead novel designs of photobioreactors with improvementsof illumination methods for high level of astaxanthm production. The maximum astaxanthin concentrations as wellas the astaxanthin yield per supplied photon were increased by at least 20% when blue or red LEDs were supplied.

• Journal of Microbiology and Biotechnology
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• 제28권12호
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• pp.2019-2028
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• 2018

Natural astaxanthin mainly derives from a microalgae producer, Haematococcus pluvialis. The induction of nitrogen starvation and high light intensity is particularly significant for boosting astaxanthin production. However, the different responses to light intensity and nitrogen starvation needed to be analyzed for biomass growth and astaxanthin accumulation. The results showed that the highest level of astaxanthin production was achieved in nitrogen starvation, and was 1.64 times higher than the control group at 11 days. With regard to the optimization of light intensity utilization, it was at $200{\mu}mo/m^2/s$ under nitrogen starvation that the highest astaxanthin productivity per light intensity was achieved. In addition, both high light intensity and a nitrogen source had significant effects on multiple indicators. For example, high light intensity had a greater significant effect than a nitrogen source on biomass dry weight, astaxanthin yield and astaxanthin productivity; in contrast, nitrogen starvation was more beneficial for enhancing astaxanthin content per dry weight biomass. The data indicate that high light intensity synergizes with nitrogen starvation to stimulate the biosynthesis of astaxanthin.

• Journal of Microbiology and Biotechnology
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• 제20권9호
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• pp.1276-1282
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• 2010

In traditional mixotrophic cultures of microalgae, all the inorganic nutrients and organic carbon sources are supplied in the medium before inoculation. In this study, however, an alternative approach was adopted in Haematococcus pluvialis Flotow, a microalga capable of growing mixotrophically on sodium acetate (Na-Ac). First, the cells were grown under 75 ${\mu}Mol$ photons $m^{-2}s^{-1}$ phototrophically without Na-Ac until the stationary phase and then exposed to five different light regimes by the addition of Na-Ac (e.g., dark, 20, 40, 75, and 150 ${\mu}Mol$ photons $m^{-2}s^{-1}$). Dry weight (DW), pigments, and especially cell number in alternative mixotrophy (AM) were higher than traditional mixotrophy (TM). Cell number in AM almost doubled up from 21.7 to $42.9{\times}10^4$ cells/ml during 5-day exposure to Na-Ac, whereas the increase was only 1.2-fold in TM. Maximum cell density was reached in 75 ${\mu}Mol$ photons $m^{-2}s^{-1}$ among the light intensities tested. We propose that Na-Ac in TM of H. pluvialis can not be utilized as efficiently as in AM. With this respect, AM has several advantages against TM such as a much higher cell density in a batch culture period and minimized risk of contamination owing to the shorter exposure of cells to organic carbon sources. In consequence, this method may be used for other strains of the species, and even for the other microalgal species able to grow mixotrophically.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2005년도 생물공학의 동향(XVII)
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• pp.216-217
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• 2005

• Korean Chemical Engineering Research
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• 제56권3호
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• pp.376-380
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• 2018

강력한 항산화물질인 astaxanthin의 함량이 다른 천연 공급원에 비해 높아 astaxanthin 생산균주로 주목받고 있는 Haematococcus pluvialis는 상당한 두께의 견고한 세포벽을 가지고 있어, 세포 파쇄를 위해 많은 에너지가 소모되고 비용이 비싼 방법들이 이용되고 있다. 이에 H. pluvialis로부터 막자와 막자사발을 이용하여 astaxanthin을 손쉽게 효율적으로 추출하는 방법을 제시하였다. 막자와 막자사발을 이용하여 분쇄한 후 추출용매로 acetonitrile, acetone, methanol, dichloromethane : methanol (1:3, v/v), ethylacetate : ethanol (1:1, v/v)로 사용하여 비교하였을 때, acetone을 이용하였을 때 astaxanthin을 1.13~1.29 배 더 높은 효율로 추출할 수 있었다. 또한 acetone으로 H. pluvialis로부터 추출할 경우, 1차 추출로 H. pluvialis에 축적되어 있는 전체 astaxanthin의 96.7%를 회수할 수 있을 정도로 acetone은 astaxanthin 추출효율이 높았다. H. pluvialis가 세포내에 축적하는 astaxanthin은 축적 특성상 ester-형태의 astaxanthin로 다량 축적하므로, 추출물 내의 다양한 형태의 astaxanthin을 분리하기 위하여 농도 구배 시스템을 적용한 HPLC 분석을 수행하였다. H. pluvialis에 축적되어 있는 전체 astaxanthin 중 free astaxanthin이 45.9%이고, 나머지 54.1%는 ester-형태의 astaxanthin이었다.

• Journal of Microbiology and Biotechnology
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• 제16권8호
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• pp.1222-1228
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• 2006

Two kinds of Haematococcus pluvialis cells (green vegetative cells cultivated under optimal cell culture conditions and red cyst cells maintained under high light stress conditions to induce astaxanthin production) were used to investigate the protein expression profiles by two-dimensional electrophoresis, image analysis, and peptide mass fingerprinting. The cellular accumulation of astaxanthin was evident after exposure to high light intensity and reached the maximum cellular level after 78 h of high light stress. In a 2-D electrophoresis analysis, 22 proteins were upregulated over 2-fold in the red cyst cells when compared with the green vegetative cells and selected for further analysis by chemically assisted fragmentation (CAF)-MALDI-TOF sequencing to identify the protein functions. Among 22 different spots, several key enzymes specific to the carotenoid pathway, including isopentenyl pyrophosphate isomerase (IPP) and lycopene $\beta$-cyclase, appeared in H. pluvialis after exposure to high light intensity. Therefore, IPP and lycopene $\beta$-cyclase would appear to be involved with carotenoid accumulation in the cytoplasm, as these peptides were preferentially upregulated by high light intensity preceding an increase in carotenoid, and only these forms were detected in the red cyst cells.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2002년도 생물공학의 동향 (X)
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• pp.220-223
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• 2002

본 연구에서는 자체 제작한 2L 규모의 bubble-column photobioreactor에서 H. pluvialis의 배양을 시도하였고 astaxanthin의 축적량 증가를 유도하기 위하여 배양시작 20일 후에 light stress를 주었다. 이 방법을 통하여 대조구에 비하여 68%의 건조균체중량 증가와 215%의 astaxanthin 축적량 증가를 유도할 수가 있었고, bubble-column photobioreactor를 이용한 유도배양이 H. pluvialis의 배양에 적합함을 알 수가 있었다.

• 한국수소및신에너지학회논문집
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• 제24권3호
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• pp.255-263
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• 2013

Carbon dioxide reduction technologies using photosynthetic microorganism were suggested to overcome environmental destruction caused by $CO_2$ in flue gases from power plant and other industries. However, there are many toxic constituents in flue gas including CO, NOx, SOx. Continuous and Excessive supply of these noxious gases to cells will leads to inhibition of microalgal growth along with partial cell death. In this study, we tested the noxious effect of SOx and NOx on the pH and microalgal growth under photoautotrophic culture in three microalgae of Neochloris oleoabundans, Chlorella vulgaris and Haematococcus pluvialis. As a result, SOx concentration more than 50 ppm led to the rapid reduction of pH, thereby inhibiting of the growth in Neochloris oleoabundans and Chlorella vulgaris. NOx concentration more the 100 ppm reduced the exponential growth of N. oleoabundans and C. vulgaris. And H. pluvialis exhibited low sensitivity to SOx and NOx. Consequently, the three microalgae of N. oleabundas, C. vulagaris and H. pluvialis showed the normal vegetative growth in 25 ppm of NOx and SOx. Above all, H. pluvialis was useful for the $CO_2$ sequestration of the flue gas including high concentrations of NOx and SOx.

• Journal of Microbiology and Biotechnology
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• 제16권12호
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• pp.1890-1896
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• 2006

Lumostatic operation for cultivation of Haematococcus pluvialis was assessed to test the scale-up strategy of photobioreactors. Lumostatic operation is a method of maintaining a proper light condition based on the specific light uptake rate ($q_e$), by cells. Lumostatic operations were performed in 0.4-, 2-, 10-, and 30-1 scale bubble column photobioreactors and the results were compared with cultures illuminated with constant light intensity. Significant differences were observed in the maximal cell concentrations obtained from 0.4-, 2-, 10-, and 30-1 scale photobioreactors under constant light intensity, yielding the maximal cell concentrations of $2.8{\times}10^5$, $2.2\times10^5$, $1.5\times10^5$, and $1.1\times10^5$ cells/ml, respectively. The maximal cell concentration in a 0.4-1 photobioreactor under lumostatic operation was $4.3\times10^5$ cells/ml. Furthermore, those in 2-, 10-, and 30-1 scale photobioreactors were about the same as that in the 0.4-1 photobioreactor. The results suggest that lumostatic operation with proper $q_e$ is a good strategy for increasing the cell growth of Haematococcus pluvialis compared with a constant supply of light energy. Therefore, lumostatic operation is not only an efficient way to achieve high cell density cultures with minimal power consumption in microalgal cultures but it is also a perfect parameter for the scale-up of photobioreactors.

• 한국미생물·생명공학회지
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• 제29권4호
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• pp.227-233
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• 2001

고부가가치의 천연 astaxanthin의 생산을 위한 Haematococcus pluvialis의 고농도 배양을 수립하기 위하여 세포 농도를 증가시키는 방법과 세포 내 색소를 증가시키는 방법 두 가지에 초점을 맞춰 본 연구를 수행하였다 Hongkong Medium (HKM) Modifed Bolds Basal Medium (MBBM) Modified Bristols Medium(MBM) Proteose-petone Medium (PPM) BG-11 Medium의 여러 배지를 사용하여 비교함으로서 배지 내 탄소원 질소원과 미량원소 등이 세포의 생장과 astaxan-tin 생산에 미치는 영향을 보면 HKM 은 $2.0$\times$10^{6}$ cells /mL의 세포 농도와 최고 6.14 mg/g cell의 astaxanthin content per cell 로 다른 배지 중 가장 높게 나타나므로 세포 농도를 높이기위해서 질소우너이 220 mg/L 인 HKM이 적당하고 색소 형성을 촉진시키기 위해서는 aggregation이 일어나 단위 세포당 astaxanthin 함량이 9.7 mg/g cell 로 높게 나타나는 MBBM이 월등함을 알수 있었다. H. pluvialis의 최적 배양 온도와 pH를 선정하깅 nl하여 $20^{\circ}C$ $25^{\circ}C$, 3$0^{\circ}C$ 온도와 pH 4.5 6.0 7.5 의 배양조건을 수립한 결가 pH $7.5 20^{\circ}C$와 $25^{\circ}C$에서는 세포생장과 색소 형성이 우수함을 관찰 할 수 있었고 배양에 적당한 20 $^{\circ}C$ 에서 배양하다가 $30^{\circ}C$로 24시간 배양항 heating 효과를 주면 astaxanthin 축적을 촉진시키는데 효과적임을 알수있었다. PPMso의 질소원과 proteose-peptone의 고갈시세포의 생장의특성과 astaxanthin 생산에 미치는 영향은 control 배지에서 세포 농도와 세포의 건조질량이 각각 $0.98$\times$10^{6}$ / cell /mL 와 0.2 g/L astaxanthin의 농도는 1.92 mg/L 단위 세포당 astaxanthin 농도는 9.6 mg/g cell 로 관찰되었다결론적으로 질소원과 peptone이 고갈되면 세포의 생장은 억제되나 astaxanthin의 생산은 촉진됨을 알수 있었으며 세포 생장을 촉진하는 광도 60$\mu$E/($\m^2$s)와 HKM 배지 이용의 1단계와 높은 광도와 MBBM배지를 이용한 색소 생산의 2단계 배양을 최적조건으로 수립하였다.