• Journal of Microbiology and Biotechnology
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• 제23권10호
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• pp.1413-1421
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• 2013

The marine microalga Isochrysis sphaerica is rich in the very-long-chain polyunsaturated fatty acids, including eicosapentaenoic acid (EPA, $C20:5{\omega}-3$) and docosahexaenoic acid (DHA, $C22:6{\omega}-3$) that are important to human health. Here, we report a functional characterization of a ${\Delta}4$-fatty acid desaturase gene (FAD4) from I. sphaerica. IsFAD4 contains a 1,284 bp open reading frame encoding a 427 amino acid polypeptide. The deduced amino sequence comprises three conserved histidine motifs and a cytochrome b5 domain at its N-terminus. Phylogenetic analysis indicated that IsFad4 formed a unique Isochrysis clade distinct from the counterparts of other eukaryotes. Heterologous expression of IsFAD4 in Pichia pastoris showed that IsFad4 was able to desaturate docosapentaenoic acid (DPA) to form DHA, and the rate of converting DPA to DHA was 79.8%. These results throw light on the potential industrial production of specific polyunsaturated fatty acids through IsFAD4 transgenic yeast or oil crops.

• 생명과학회지
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• 제20권4호
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• pp.561-571
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• 2010

대장암은 미국 등 서양 국가뿐만 아니라 국내에서도 2번째로 많이 발병이 되는 암으로 알려져 있다. 역학조사에 의하면 ${\omega}3$-PUFAs를 많이 섭취한 인종에서 대장암 발생빈도가 감소하고 최근 ${\omega}3$-PUFAs는 수종의 암에 대해 항암작용을 나타낸다고 한다. 이에 본 연구에서는 대장암에서 DHA 등 ${\omega}3$-PUFA의 항침윤 기전을 규명하여 다음과 같은 결과를 얻었다. DHA및 EPA는 대장암 세포주 SW480의 증식을 농도 의존적으로 억제하였으나 AA는 거의 영향이 없었으며 TUNEL assay로 apoptotic cell death가 확인 되었다. DHA는 $\beta$-catenin 단백 및 TCF/LEF luciferase 활성을 농도 의존적으로 억제 하였다. SW480 세포의 침윤능은 DHA의 농도에 의존적으로 억제되었다. DHA처리 후 MMP-9 및 MMP-2 mRNA양이 감소되었을 뿐만 아니라 그 promoter의 reporter 활성도 억제되었다. NF-kB 및 p-IkB 단백질양도 DHA의 처리농도에 의존적으로 감소하였으며 NF-kB promoter의 활성도 억제되었다. 이상의 결과로 ${\omega}3$-PUFA는 대장암에서 NF-kB 신호전달 차단에 의한 MMP-2 및 MMP-9 발현을 억제하여 침윤을 억제하여 항암작용을 나타낼 수 있음을 시사하며, 따라서 ${\omega}3$-PUFA는 대장암의 예방 및 치료에 유용하게 사용될 수 있으리라 생각된다.

• 생명과학회지
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• 제23권5호
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• pp.689-697
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• 2013

오메가-3 지방산은 많은 암에서 세포독성을 나타낸다고 보고 되어 왔으나 구강암에 대한 연구는 전혀 없다. 이에 본 연구에서는 구강암세포에서 오메가-3 지방산 중 DHA의 세포독성 기전을 규명하여 다음과 같은 결과를 얻었다. DHA는 구강암 세포주 SCC-4 및 SCC-9의 증식을 농도 의존적으로 억제하였으며, FACS 분석, TUNEL assay 및 PARP cleavage 등에 의해 자가사멸을 유도함이 확인 되었다. 또한 DHA는 LC-3II 단백증가, GFP-LC-3 dot 형성 및 autophagic flux assay 등에 의해 자가포식도 유도됨이 규명되었다. SCC-9 세포에서 AMPK의 인산화는 DHA 에 의해 증가 하였으나, p-$AKT^{Thr308}$, p-$AKT^{Ser473}$ 및 mTOR단백양은 감소하였다. 이상의 결과로 DHA는 구강암세포에서 AMPK 활성증가 및 AKT 억제에 통한 mTOR 신호경로 차단에 따른 자가사멸 및 자가포식에 의해 세포독성을 나타낼 수 있음을 시사하며, 따라서 DHA는 구강암의 예방 및 치료에 유용하게 사용될 수 있으리라 생각된다.

• Asian-Australasian Journal of Animal Sciences
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• 제30권7호
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• pp.973-978
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• 2017

Objective: The aim of this study was to investigate the effects of simultaneous supplementation of laying hens with alpha-linolenic acid (ALA) resources (flax, perilla, and Eucommia ulmoides [E. ulmoides] seeds) and eicosapentaenoic acid/docosahexaenoic acid (EPA/DHA) resources (Schizochytrium sp.) on egg quality and n-3 polyunsaturated fatty acids (PUFA) profile. Methods: Dietary treatments were as follows: i) diet C (control diet); ii) diet F (diet C+10% flaxseeds); iii) diet P, (diet C+10% perilla seeds); iv) diet E (diet C+10% E. ulmoides seeds); v) diet A (diet C+1.5% microalage); vi) diet AF (diet C+10% flaxseeds+1.5% microalage); vii) diet AP (diet C+10% perilla seeds+1.5% microalgae); viii) diet AE (diet C+10% E. ulmoides seeds+ 1.5% microalage). Results: Egg weight, yolk weight and production ratio were not significantly affected by either algae or in combination with seeds (p>0.05). No significant difference was observed in ALA and DHA concentration in eggs between flaxseed, perila, and E. ulmodies seeds supplementation alone (p>0.05). N-3 PUFA in eggs was slightly improved by microalgae supplementation. The best supplementation, a combination of microalgae and perilla seeds, elevated (p<0.05) ALA from 19.7 to 202.5 mg/egg and EPA+DHA from 27.5 to 159.7 mg/egg. Highest n-3 PUFA enrichment (379.6 mg/yolk) was observed with supplementation of a combination of perilla seed and microalgae (362.2 mg/yolk), followed by a combination of flaxseed and microalgae (348.4 mg/yolk). The ALA, EPA, and DHA content obtained with a combination of microalgae and seeds surpassed the total sum of that obtained with microalgae or ALA-seeds alone. Conclusion: It is feasible to enrich eggs with n-3 PUFAs by perilla or E. ulmodies seeds instead of flaxseeds. Simultaneous supplementation of microalgae and seeds helped improve the transfer from EPA and docosapentaenoic acid into DHA.

• 약학회지
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• 제38권1호
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• pp.57-66
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• 1994

We studied the effect of eicosanoids on the content of energy metabolites and the intrasynaptosomal $Ca^{++}([Ca^{++}]_i)$ concentration in cerebral ischemic rats. An ischemic model was made by bilateral carotid artery ligation (BCAL) and by incubation of synaptosomes under aglycemic and $N_2$ gas bubbling condition. The content of ATP, creatine phosphate and glucose decreased at 15 minutes after BCAL while that of lactate increased in male Wistar rats. Oral administration of EPA(100 mg/ml/Kg/day) or DHA(16 mg/ml/Kg/day) for 6 weeks improved both the decreases and the increase of the cerebral energy metabolites. In addition, the increase of $[Ca^{++}]_i$, under BACL was suppressed by EPA or DHA treatment. When the both Wistar rats and SHR were administered orally with EPA or DHA for 6 weeks, the effect on the increase of $[Ca^{++}]_i$ under ischemia by $N_2$ gas bubbling were protected. From these results, it may be that EPA or DHA treatment were greatly contributed to preservation of ischemic cerebral energy metabolism and $Ca^{++}$ concentration.

• 한국수산과학회지
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• 제51권5호
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• pp.549-555
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• 2018

The objectives of this study were to confirm the nutritional requirements and improve the survival of the walleye pollock Theragra chalcogramma, a cold seawater fish, by enrichment. We analyzed the fatty acids and amino acids of fertilized pollock eggs before hatching, just-hatched larvae, larvae that had absorbed only the yolk sac, and larvae starved for 2 days after yolk absorption. For the survival improvement experiment, we administered docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), and DHA-EPA enrichment. Fatty acid decreased DHA and EPA content. On the $30^{th}$ day, body growth was significantly greater in pollock given the EPA and DHA-EPA treatments (P<0.05). Larval survival at 20 and 30 days after hatching (DAH) was greatest under the DHA-EPA treatment (P<0.05). Survival was significantly lower under the EPA treatment at 10 DAH, but then increased to approach that seen under the EPA-DHA treatment (P<0.05). Therefore, we determined that reduced survival in hatchlings of high-mortality pollack could be improved by controlling EPA and DHA content during enrichment. We conclude that cold seawater fish must be given feed that meets their nutritional needs, which can be accomplished using newly manufactured enrichment products for the larvae of cold seawater fish, such as pollock.

• 한국식품과학회지
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• 제22권5호
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• pp.520-525
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• 1990

포화지방산 또는 불포화지방산의 분리방법인 요소부 가법이 지닌 가메탄올 분해(methanolysis)와 농축물 회수의 곤란함 등의 문제를 해결하기 위하여 지방산과 요소의 부가화합물 형성의 반응매질로 유기용매를 사용하고 요소의 습윤제로 물을 사용한 새로운 요소부가법을 제시하였다. 어유를 원료로 제조한 지방산을 헵탄, 헥산, 이소옥탄같은 비극성 유기용매에 용해키시고 요소와 요소의 습윤제인 물을 첨가하여 부가화합물 형성을 유도한 후 요소와 부가화합물을 잘 형성하지 않는 불포화지방산 분획을 회수하여 지방산 조성을 분석한 결과 EPA, DHA 및 이들의 전구물질을 포함한 고도불포화지방산이 80%이상인 것으로 나타났다. 이 고도불포화지방산 농축물내에 EPA와 DHA 함량은 사용하는 유기용매으 종류에 따라서 변화되었는데, 헵탄과 에틸 에테르 등을 사용할 경우에는 EPA의 농축에 효과가 높았다. 이러한 특성을 이용하여 우선 펜탄을 사용하여 DHA를 농축하고 다시 헵탄을 사용하여 EPA의 농축을 시도한 바 어유 지방산으로부터 DHA가 50%인 농축물과 EPA가 53%인 분획을 각각 얻을 수 있었다.

• 청정기술
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• 제17권3호
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• pp.266-272
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• 2011

초임계 이산화탄소 및 유기용매를 이용하여 동결 건조된 멸치 시료로부터 오일을 추출하였으며, 시료는 700 ${\mu}m$로 균질화 시켜 사용하였다. 다양한 압력 (15~25 MPa) 및 온도 ($40{\sim}60^{\circ}C$) 조건에서 실험을 수행하였으며, 22 g/min의 일정한 유량을 추출기로 유입시켰다. 추출된 오일의 지방산 조성을 확인하기 위하여 가스 크로마토그래피를 이용하여 분석한 결과 추출된 멸치 오일에는 myristic acid, palmitic acid, stearic acid, palmitoleic acid, eicosapentaenoic acid (EPA), 그리고 docosahexaenoic acid (DHA)가 주요 지방산으로 확인되었다. 추출된 오일 내 EPA 및 DHA와 같은 고도불포화지방산(PUFAs)함량을 비교하였을 때, 유기용매 추출 오일보다 초임계 이산화탄소를 이용하여 추출한 오일의 고도 불포화 지방산 함량이 더 높았다. 산화도 비교 실험인 산가 및 과산화물가 값을 측정한 결과, 유기용매 추출 오일보다 초임계 이산화탄소를 이용한 추출 오일의 산화도가 유의적으로 낮았으며, 산화되는 속도 또한 느리게 진행되는 것을 확인할 수 있었다.

• Journal of Nutrition and Health
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• 제30권8호
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• pp.987-994
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• 1997

To investigate the effects of n-3 fatty acids on breast cancer, MDA-MB231 human breast cancer cells were cultured in the presence of $\alpha$-linolenic (LNA), eicosapentaenoic(EPA), and docosahexaenoic acid (DHA) at a concentration of 0.5$\mu\textrm{g}$/ml in serum -free IMM medium. Cell growth was monitored and thiobarbituric acid reactive substances (TBARS), $\alpha$-tocopherol contents, and oncogene expression were measured. To compare the effects of n-3 fatty acids with other types of fatty acid, steraic (STA), olieic(OA). linoleic acid(LA) were used. After one day , cell growth was retarded most highly when DHA was in the medium. Cellular TBARS level measured after three days of culture was the highest with DHA in the medium and was also increased by LNA and EPA, compared with STA, OA and LA. Alpha-tocoopherol contents of cells were decreased by DHA but only modestly. There was non significant difference in $\alpha$-tocopherol contents in cells cultured in the presence of the other fatty acids. northern blot hybridization carried out with cells cultured during 24 hours showed that levels of erbB-2 mRNA were not altered by six different fatty acids in the medium but those of c-myc were transiently decreased in the early period by both n-6 and n-3 polyunsaturated fatty acids. The level of tumor suppressor gen p53 mRNA , however, was increased by DHA with time. It is concluded that the cytotoxicity of lipid peroxide and increased expression of tumor suppressor gene p53 are at least partly responsible for the inhibitory effect of DHA on growth of breast cancer cells.

• Biotechnology and Bioprocess Engineering:BBE
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• 제7권1호
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• pp.10-15
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• 2002

Environmental and medium factors were investigated as basic data for optimizing DHA production when using Thraustochytrium aureum. To study the effect of environmental conditions, the rotation speed and culture temperature were changed. Plus the trend of the growth characteristics, lipid content in the biomass, and DHA content in lipids were evaluated according to various initial glucose concentrations. The biomass, lipid, and DHA analyses showed that the physiological characteristics of T. aureum were closely related with the environmental and medium conditions, as in the case of other marine microorganisms. For example, a low rotation speed of 50 rpm lowered the cell growth rate as well as the DHA content in the lipids. A low temperature had a negative effect on the cell growth, yet a positive effect on the lipid content in the biomass. Different initial glucose concentrations had no effect on the lipid content in the biomass or DHA content in the lipids, yet did affect the cell growth. Accordingly, these results show that environmental and medium factors must be synthetically considered in order to optimize DHA production when using T. aureum.