• Journal of Microbiology and Biotechnology
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• 제26권5호
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• pp.854-866
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• 2016

The production cost of biodiesel from microalgae is still not competitive, compared with that of petroleum fuels. The genetic improvement of microalgal strains to increase triacylglycerol (TAG) accumulation is one way to reduce production costs. One of the most promising approaches is the isolation of starch-deficient mutants, which have been reported to successfully increase TAG yields. To date, such a stable mutant is not available in an oleaginous marine microalga, despite several advantages of using marine species for biodiesel production. Algae in the genus Dunaliella are known to tolerate high salt concentration and other environmental stresses. In addition, the cultivation processes for large-scale outdoor commercialization have been well established for this genus. In this study, Dunaliella tertiolecta was used to screen for starch-deficient mutants, using an iodine vapor-staining method. Four out of 20,016 UV-mutagenized strains showed a substantial reduction of starch content. A significantly higher TAG content, up to 3-fold of the wild-type level, was observed in three of the mutants upon induction by nitrogen depletion. The carotenoid production and growth characteristics of these mutants, under both normal and oxidative stress conditions, were not compromised, suggesting that these processes are not necessarily affected by starch deficiency. The results from this work open up new possibilities for exploring Dunaliella for biodiesel production.

• 수산해양교육연구
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• 제28권2호
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• pp.315-322
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• 2016

This study determined the optimum microalgae species and concentration for growth of small rotifer Proales similis, in community cultures by feeding it several different foods. Four single trials (Tetraselmis suecica, TET; Isochrysis galbana, ISO; Nannochloropsis oculata, NAN; freshwater Chlorella vulgaris, CHL) were conducted at 5 psu and $25^{\circ}C$. The maximum density and specific growth rate (SRG) were highest for rotifer fed CHL reaching 1,075 individuals (inds.)/mL and 0.83 respectively. By contrast, other foods of ISO, NAN and TET resulted in poor growth rates and maximum density. In the freshwater C. vulgaris experiments examining a range of 0.05 to $8.8mg/10^4rotifers$, the growth of rotifer tended to decrease with the amount of supplement. The maximum density and SGR of females was highest at 910 inds./mL and 0.55 respectively, at $0.05mg/10^4rotifers$. These results suggest that the best microalgae species for the culture of P. similis is freshwater C. vulgaris and the optimum concentration is $0.05mg/10^4rotifers$.

• 한국해양바이오학회지
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• 제16권1호
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• pp.45-54
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• 2024

Microalgae have great potential in the biomedical and pharmaceutical industries as a new type of bioreactor that can produce proteins for specific purposes, including recombinant proteins, pharmaceuticals, and industrial enzymes. Despite the production advantages and importance of microalgae-based expression systems, studies on secretion efficiency are limited. In this study, for stable expression and efficient secretion of the heterologous protein (human SCF and human INFγ) in Chlorella vulgaris, we constructed SP:hSCF:His and SP:hINFγ:His plant binary vectors using the signal peptide (SP) of Chlamydomonas reinhardtii, and we obtained stable transformants through the effective agrobacterium-mediated transformation of these vectors. Transformants with accurately inserted hSCF and hINFγ demonstrated stably increased mRNA and protein expression using RT-PCR and western blotting under the same culture conditions. Following the analysis of the proteins secreted into the culture medium using ELISA, it was confirmed that hINFγ was effectively produced in the transformed C. vulgaris culture medium. The overall findings indicate that the combination of heterologous protein and SP may be crucial for ensuring the expression and secretion of recombinant proteins in Chlorella culture systems.

• Journal of Microbiology and Biotechnology
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• 제28권2호
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• pp.267-274
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• 2018

Lipids in microalgae are energy-rich compounds and considered as an attractive feedstock for biodiesel production. To redirect carbon flux from competing pathways to the fatty acid synthesis pathway of Tetraselmis sp., we used three types of chemical inhibitors that can block the starch synthesis pathway or photorespiration, under nitrogen-sufficient and nitrogen-deficient conditions. The starch synthesis pathway in chloroplasts and the cytosol can be inhibited by 3-(3,4-dichlorophenyl)-1,1-dimethylurea and 1,2-cyclohexane diamine tetraacetic acid (CDTA), respectively. Degradation of glycine into ammonia during photorespiration was blocked by aminooxyacetate (AOA) to maintain biomass concentration. Inhibition of starch synthesis pathways in the cytosol by CDTA increased fatty acid productivity by 27% under nitrogen deficiency, whereas the blocking of photorespiration in mitochondria by AOA was increased by 35% under nitrogen-sufficient conditions. The results of this study indicate that blocking starch or photorespiration pathways may redirect the carbon flux to fatty acid synthesis.

• Fisheries and Aquatic Sciences
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• 제11권4호
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• pp.205-208
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• 2008

The mixotrophic dinoflagellate Cochlodinium polykrikoides was reported to be linked to major fish kills in Korea and Japan since the 1990s. Rapid and sensitive detection of microalgae has been problematic because morphological identification of dinoflagellates requires light microscopic and scanning electron microscopic observations that are time consuming and laborious compared to real-time PCR. To address this issue, a real-time PCR probe targeting the ITS2 rRNA gene was used for rapid detection and quantification of C. polykrikoides. PCR inhibitors in water column samples were removed by dilution of template DNA for elimination of false-negative reactions. A strong association between cell quantification using real-time PCR and microscopic counts suggests that the real-time PCR assay is an alternative method for cell estimation of C. polykrikoides in environment samples.

• Journal of Industrial and Engineering Chemistry
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• 제68권
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• pp.267-273
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• 2018

Hydrothermal liquefaction of Chlorella vulgaris feedstock containing 80% (w/w) water was conducted in a batch reactor as a function of temperature (300, 325 and $350^{\circ}C$) and reaction times (5, 10 and 30 min). The biocrude yield, elemental composition and higher heating value obtained for various reaction conditions helped to predict the optimum conditions for maximizing energy recovery. To optimize the recovery of inorganic nutrients, we further investigated the effect of reaction conditions on the ammonium ($NH_4{^+}$), phosphate ($PO_4{^{3-}}$), nitrate ($NO_3{^-}$) and nitrite ($NO_2{^-}$) concentrations in the aqueous phase. A maximum energy recovery of 78% was obtained at $350^{\circ}C$ and 5 min, with a high energy density of 34.3 MJ/kg and lower contents of oxygen. For the recovery of inorganic nutrients, shorter reaction times achieved higher phosphorus recovery, with maximum recovery being 53% at $350^{\circ}C$ and 5 min. Our results indicate that the reaction condition of $350^{\circ}C$ for 5 min was optimal for maximizing energy recovery with improved quality, at the same time achieving a high phosphorus recovery.

• 한국해양바이오학회지
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• 제2권3호
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• pp.174-186
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• 2007

Amphora coffeaeformis and Achnanthes longipes are commonly found as dominant benthic microalgae in Jeju coastal water throughout the year. In order to investigate pharmaceutical uses of these diatoms, each single species was isolated with micropipette under phase contrast microscope and subcultured with synthetic seawater media which was enriched with F/2 media, trace metal solution and $Na_2SiO_3$). Growth characteristics of these species were also determined with different combination of salinity, nutrients concentration and temperature. Thereafter, mass culture of each species was done based on the maximum growth condition. Biomass was collected after two weeks of mass culture and freeze dried for antioxidant study. The antioxidant properties of different fractions (n-hexane, chloroform and ethylacetate) obtained by solvent fractionation of 80% methanolic extract of two microalgae were investigated for free radical, reactive oxygen species scavenging (Super oxide, Hydrogen peroxide, Hydroxyl radical and Nitric oxide), metal chelating and lipid peroxidation inhibition activities. All fractions of A. longipes showed higher $DPPH^{\cdot}$ (free radical) scavenging activities (n-hexane: 89.0%, Chloroform: 76.0%, Ethylacetate: 66.0%, Methanol: 90.6% and aqueous residue: 63.0%). N-hexane fraction of A. longipes showed significantly higher activity (49.0%) on nitric-oxide. Ethylacetate fraction of A. longipes and aqueous residue of A. coffeaeformis exhibited 64.0% and 75.6% metal chelating activity which was higher than commercial antioxidants (${\alpha}$-tocopherol: 18.0% and BHT: 16.0%). The n-hexane fraction of A. coffeaeformis had 67.5% activity on $DPPH^{\cdot}$. Chloroform and n-hexane fractions of A. coffeaeformis exhibited 46.2% and 47.6% $H_2O_2$ scavenging effects which were closely similar to commercial antioxidants (${\alpha}$-tocopherol: 49.2% and BHT: 58.6%). Chloroform and ethylacetate fractions of A. longipes and fraction of n-hexane and chloroform of A. coffeaeformis showed better lipid peroxidation activities than ${\alpha}$-tocopherol. These data suggest that both organic and aqueous fractions have good antioxidative compounds with different antioxidant properties.

• 한국미생물·생명공학회지
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• 제48권3호
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• pp.328-336
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• 2020

본 연구에서는 국내 낙동강 수계에서 신규하게 분리된 미세조류인 Parachlorella sp. 종의 바이오매스 및 지방산 생산성에 대한 배지의 영향을 연구하였다. 미세조류 배양에 통상적으로 사용되는 BG-11, TAP, BBM 배지를 사용하여 바이오매스 생산성은 TAP 배지에서, 지방산 축적은 BBM 배지에서 가장 잘 일어나는 것으로 확인되었고, 지방산 생산성을 향상시키기 위해 암모니아와 아세트산을 사용하는 TAP 배지의 조성을 변화하여 BBM 배지처럼 지방산 축적을 유도하며 바이오매스 생산성을 증가시킨 MTAP 배지를 개발하였다. 전체적인 바이오매스와 지방산 생산성을 높이기 위해서는 MTAP-1 배지가 적합하여 바이오매스 생산성과 지방산 생산성은 기존의 TAP 배지 대비 각각 14%, 45% 증가하였다. 생리 활성 효과로 인해 관심도가 높은 오메가-3 지방산의 생산에는 MTAP-4 배지가 가장 적합하여 바이오매스 생산성과 오메가-3 지방산 생산성이 기존 BBM 배지 대비 각각 18%, 39% 증가하여 목표 중점 생산물질(바이오매스, 총 지방산, 또는 오메가-3 지방산)의 생산성을 향상시킬 수 있는 신규 배지 2종의 조성을 개발하였다.

• 한국식품위생안전성학회지
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• 제35권5호
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• pp.411-418
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• 2020

본 연구에서는 제외국 관리 해양생물독소인 YTXs의 원인조류, 독화생물, 국내·외 관리현황 및 분석법에 대해 리뷰하였다. 현재까지 YTXs에 의한 국내 수산물 오염이 보고된 사례는 없지만 기후변화로 인한 국내 연안 해역의 아열대화로 독성조류의 국내 유입 가능성이 있을 뿐 아니라 외국의 경우 YTXs에 의한 오염이 빈번하게 발생하고 있기 때문에 해당 국가로부터 수입되는 수산물의 경우 잠재적인 오염가능성이 있어 이에 대한 선제적 대응이 필요하다. YTXs는 와편모조류인 Protecratium reticulatum, Gonyaulax polygramma, Gonyaulax spinifera과 Lingulodinium polyedrum에 의해 생성되는 것으로 알려져 있으며, 이들은 국내 연안에도 출현한다. 현재 패류 내 YTXs는 메탄올 추출, SPE 카트리지 정제 후 LC-MS/MS를 이용하여 분석하고 있다. YTXs, pectenotoxins, azaspiracids와 같은 지용성 패독은 추출 및 정제 등의 전처리 과정이 유사하여 동시분석이 가능할 것으로 보이며, 이에 대한 분석법 최적화가 필요하다. 향후 국내 해역의 YTXs의 원인조류 출현 및 국내 유통(국내산 및 수입산) 수산물의 오염에 대한 지속적인 모니터링이 필요하다.

• 한국수산과학회지
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• 제40권3호
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• pp.122-127
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• 2007

Photosensitized extracts of 28 seaweed species were tested in vitro to examine the variation in their algicidal activities against the toxic microalga Cochlodinium polykrikoides. When both seaweed extracts and microalgae were exposed to UV and visible light, methanol extracts of Porphyra yezoensis showed 5.7- and 4.4-fold increasesin light-dependent algicidal activity, respectively. When only seaweed extracts were exposed to both UV (365 nm) and visible light (white lamp) at the same time, methanol extracts of Enteromorpha linza and Carpopeltis affinis showed 3.3- and 3.4-fold increases in algicidal activity, respectively. When UV-photosensitized extracts were left in the dark, the algicidal activity of Ecklonia cava increased 13-fold after 5 h. When visible light-photosensitized extracts were left in the dark, the algicidal activity of Monostroma nitidum increased by 3.3-fold in 1 h.