LED 광원에 따른 미세조류 Haematococcus pluvialis의 성장 특성

이건우,김송이,유용진,이영복,김진우,김호섭 한국산학기술학회 2020 한국산학기술학회논문지 Vol.21 No.10

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. 본 연구에서는 미세조류 Haematococcus pluvialis의 성장에 미치는 배양 배지와 광원의 영향을 평가하기 위해 제한 성분 배지인 Modified Bold’s Basal Medium (MBBM) 와 상업용 액체 비료 배지인 Neo, 그리고 파장이 서로 다른 7개의 광원을 사용하여 H. pluvialis를 39일 동안 배양하고 성장률을 비교하였다. 그 결과 제한 성분 배지인 MBBM에서 H. pluvialis의 성장은 fluorescent light 광원에서 세포 성장이 가장 높게 나타난 반면, blue+red LED에서 세포 성장이 가장 낮게 나타났다. 상업용 배지인 Neo에서 H. pluvialis의 성장은 fluorescent light 광원에서 세포 성장이 가장 높게 나타났으며, blue LED에서는 세포 성장이 가장 낮음이 확인되었다. 본 연구를 통해 MBBM 배지가 Neo 배지보다 우수한 결과를 나타내었으며 또한, MBBM 배지를 이용하여 fluorescent light에서 성장한 미세조류는 본 연구에서 가장 우수한 세포 성장 결과를 보였다. 이러한 결과는 이차대사산물 생산을 위한 H. pluvialis 배양에서 배지, 광원과 광량을 최적화한 것으로 미세조류 대량 배양에 기초 자료로써 유용하게 활용될 것이다.

Enhancement of microalga <i>Haematococcus pluvialis</i> growth and astaxanthin production by electrical treatment

Kim, Jee Young,Lee, Changsu,Jeon, Min Seo,Park, Jaewon,Choi, Yoon-E Elsevier 2018 Bioresource technology Vol.268 No.-

<P><B>Abstract</B></P> <P>In this study, we investigated the effects of electrical treatment on <I>Haematococcus pluvialis</I> growth. The slow growth of <I>H. pluvialis</I> is a major limitation for its mass production. We discovered that electrical treatment may promote the growth of <I>H. pluvialis</I>. To evaluate optimal growth-promoting conditions, the algal growth rate was investigated at various voltages. The optimum current was identified as 100 mA (voltage: 25 V). In comparison with the non-treated cells, those subjected to electrical treatment showed a 1.2 fold increase in cell density. Further experiments confirmed the direct impact of electrical treatment on the growth of <I>H. pluvialis</I>. The periodic application of electrical voltage resulted in a significant increase in the dry weight and astaxanthin production. The astaxanthin content in the periodic application of electrical treatment was 32.6 mg/L, which was a 10% increase compared to those in the non-treated controls. This strategy may serve as a novel approach to enhance <I>H. pluvialis</I> growth as well as astaxanthin production.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Electrical treatment was deployed to enhance the growth of microalga <I>Haematococcus</I>. </LI> <LI> Direct electrical treatment of <I>H. pluvialis</I> cells resulted in growth-promoting effects. </LI> <LI> Periodic electrical treatment was shown to be more effective. </LI> <LI> Our results provide a foundation to increase the productivity of <I>H. pluvialis</I>. </LI> </UL> </P>

초음파처리를 이용한 Haematococcus pluvialis로부터의 아스타잔틴의 추출 및 분석

김소영(Soyoung Kim),조은아(Eunah Cho),유지민(Jimin Yoo),인만진(Man-Jin In),채희정(Hee Jeong Chae) 한국식품영양과학회 2008 한국식품영양과학회지 Vol.37 No.10

본 연구에서는 헤마토코커스 플루비알리스(H. pluvialis)의 파쇄세포(cracked cell)로부터 추출된 아스타잔틴의 HPLC 분석방법을 확립하고, UV/visible 및 FT-IR 분광분석법을 이용하여 헤마토코커스 추출물과 합성 아스타잔틴의 구조적 특성을 조사하였다. 아스타잔틴을 메탄올 용매에 녹이고, 45분 정도 초음파로 처리할 경우 교반처리하는 경우보다 1.5배 정도 용해율이 높아지는 것을 확인하였다. 에스터 형태로 존재하는 추출물 중의 아스타잔틴의 분석을 위해 cholesterol esterase를 이용해 헤마토코커스 추출물을 가수 분해처리를 하여 유리형으로 전환시킨 후 HPLC 분석을 수행하였다. 아스타잔틴의 함량을 분석하기 위해 역상칼럼(C18)과 UV/visible 검출기를 사용하였고, 이동상은 메탄올과 물(95:5)의 혼합용매를 사용하여 분석하였다. 효소처리 후, 헤마토코커스 추출물의 흡수스펙트럼이 합성 아스타잔틴과 유사한 패턴으로 변화하는 것을 확인함으로써 헤마토 코커스로부터의 아스타잔틴의 추출 및 분석조건을 확립하였다. 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 hydrolysate was changed to similar pattern to synthetic astaxanthin, confirming the extraction and analysis condition of astaxanthin from H. pluvialis.

미세조류로부터의 에너지 효율적인 Astaxanthin 회수 기술 개발

김선영 ( Sun Young Kim ),오유관 ( You-kwan Oh ),하성호 ( Sung Ho Ha ) 한국화학공학회 2018 Korean Chemical Engineering Research(HWAHAK KONGHA Vol.56 No.3

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. The astaxanthin recovery efficiencies were compared in acetonitrile, acetone, methanol, dichloromethane : methanol (1:3, v/v) and ethylacetate : ethanol (1:1, v/v) as a extraction solvent after the grinding of the H. pluvialis cells. The astaxanthin extraction yield in acetone was 1.13~1.29 times higher than other extraction solvents. It was also found that 96.7% of astaxanthin accumulated in H. pluvialis could be recovered by a single extraction. Since astaxanthin exists mainly as astaxanthin esters in H. pluvialis, a gradient reversed-phase HPLC analysis was carried out for the separation of astaxanthin esters from the extracts of H. pluvialis. Among the astaxanthin inside the H. pluvialis cell, free astaxanthin was 45.9% and astaxanthin esters were the rest.

Cold-tolerant strain of Haematococcus pluvialis (Haematococcaceae, Chlorophyta) from Blomstrandhalvøya (Svalbard)

Klochkova, Tatyana A.,Kwak, Min Seok,Han, Jong Won,Motomura, Taizo,Nagasato, Chikako,Kim, Gwang Hoon The Korean Society of Phycology 2013 ALGAE Vol.28 No.2

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.

Cold-tolerant strain of Haematococcus pluvialis (Haematococcaceae, Chlorophyta) from Blomstrandhalvøya (Svalbard)

Tatyana A. Klochkova,곽민석,한종원,Taizo Motomura,Chikako Nagasato,김광훈 한국조류학회I 2013 ALGAE Vol.28 No.2

A new cold-adapted Arctic strain of Haematococcus pluvialis from Blomstrandhalvøya 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°C), whilst the usual growth temperature for H. pluvialis is between 20-27°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.

Enhancing Astaxanthin Accumulation in Haematococcus pluvialis by Coupled Light Intensity and Nitrogen Starvation in Column Photobioreactors

( Wen-wen Zhang ),( Xue-fei Zhou ),( Ya-lei Zhang ),( Peng-fei Cheng ),( Rui Ma ),( Wen-long Cheng ),( Hua-qiang Chu ) 한국미생물 · 생명공학회 2018 Journal of microbiology and biotechnology Vol.28 No.12

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 μmo/m2/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.

Ultrasonic Treatment Enhanced Astaxanthin Production of Haematococcus pluvialis

Park Yun Hwan,Park Jaewon,Choi Jeong Sik,Kim Hyun Soo,Choi Jong Soon,Choi Yoon-E 한국미생물학회 2023 The journal of microbiology Vol.61 No.6

In this study, effects of ultrasonic treatment on Haematococcus pluvialis (H. pluvialis) were investigated. It has been confirmed that the ultrasonic stimulation acted as stress resources in the red cyst stage H. pluvialis cells containing astaxanthin, resulting in additional astaxanthin production. With the increase in production of astaxanthin, the average diameter of H. pluvialis cells increased accordingly. In addition, to determine how ultrasonic stimulation had an effect on the further biosynthesis of astaxanthin, genes related to astaxanthin synthesis and cellular ROS level were measured. As a result, it was confirmed that astaxanthin biosynthesis related genes and cellular ROS levels were increased, and thus ultrasonic stimulation acts as an oxidative stimulus. These results support the notion on the effect of the ultrasonic treatment, and we believe our novel approach based on the ultrasonic treatment would help to enhance the astaxanthin production from H. pluvialis.

Haematococcus pluvialis로부터 아스타잔틴 추출을 위한 추출공정 조건 최적화

김송이,전성진,김호섭,감다혜,김진우 한국생물공학회 2019 KSBB Journal Vol.34 No.4

In this study, the selection of extraction solvent and optimization of extraction condition were carried out to increase the extraction of astaxanthin, an antioxidant material produced from Haematococcus pluvialis. For increasing the extraction of astaxanthin through more effective cell disruption, the concentration of acetone, rotation speed of homogenizer, extraction temperature, and extraction time were sequentially optimized. When acetone (50% v/v) was used as an extract solvent, the extraction concentration of astaxanthin was found to be the highest (1.41 μg/mL), and the extraction concentration increased 3.3 times compared to ethanol extraction. As the concentration of acetone increased to 99.5% (v/v), it was found that the concentration of astaxanthin increased proportionally to 2.47 μg/mL. When evaluating the effect of rotation speed and extraction temperature, astaxanthin concentration was maximized to 3.93 g/mL at 15,318 rpm (impeller tip speed = 5.67 m/s) and 52.5oC. In addition, as extraction time increased, the concentration of extract was increased and resulted in a maximum of 4.76 μg/mL at 16.6 min which was a 10.9-fold increase compared to ethanol extraction indicating that acetone is more effective than conventional solvents in the extraction of astaxanthin.

Enhanced autotrophic astaxanthin production from Haematococcus pluvialis under high temperature via heat stress-driven Haber-Weiss reaction

김우현,홍민의,심상준 한국공업화학회 2015 한국공업화학회 연구논문 초록집 Vol.2015 No.1

High temperatures (30-36°C) inhibited astaxanthin accumulation in H. pluvialis under photoautotrophic conditions. The depression of carotenogenesis was primarily attributed to excess intracellular less reactive oxygen species (LROS; O₂^- and H₂O₂) levels generated under high temperature conditions. Here we show that the heat stress-driven inefficient astaxanthin production was improved by accelerating the iron-catalyzed Haber-Weiss reaction to convert LROS into more reactive oxygen species (MROS; O₂ and OHㆍ), thereby facilitating lipid peroxidation. As a result, during 18 days of photoautotrophic induction, the astaxanthin concentration of cells cultured in high temperatures in the presence of iron (450 μM) was dramatically increased by 75% (30°C) and 133% (36°C) compared to that of cells exposed to heat stress alone. The heat stress-driven Haber-Weiss reaction will be useful for economically producing astaxanthin by reducing energy cost and enhancing photoautotrophic astaxanthin production, particularly outdoors utilizing natural solar radiation including heat and light for photo-induction of H. pluvialis.