$Q_{10}$ 함유 광합성세균의 반응특성

정수경,김중균,Jeong, Soo Kyoung,Kim, Joong Kyun 한국해양바이오학회 2007 한국해양바이오학회지 Vol.2 No.3

Cultivation of a $Q_{10}$-producing photosynthetic bacterium, Rhodobacter sphaeroids, was carried out in a 1-L flask to characterize its cellular growth reaction. The result of experiment showed that dissolve oxygen in the broth was depleted within 7 h. ORP decreased with decrease of DO, and recovered somewhat with increase of pH. The growth of R. spahaeroids reached at late-log phase in 140 h of reaction. The final pH and dry-cell weight were 7.62 and 2.2 mg/mL, respectively. The $Q_{10}$ content in the final broth was 2.35 mg/g dry cell weight, which was higher than that obtained in tube culture.

3T3-L1 및 B16F10 세포에서 청각 메탄올 추출물에 의한 지방 세포 분화 및 멜라닌 생성의 억제 효과

최은옥,최영현,황혜진 (사)한국해양바이오학회 2021 한국해양바이오학회지 Vol.13 No.1

Codium fragile (Suringar) Hariot, a green alga of the Codiales family, has been reported to have several bioactive properties, including antioxidant and anti-inflammatory properties. However, its antiobesity and whitening effects and their underlying mechanisms are unclear. This study aimed to evaluate the antiobesity and melanogenesis inhibitory effects of C. fragile using methanol extracts of C. fragile (MECF). The results of this study revealed that MECF inhibited the accumulation of lipid droplets and triacylglycerol in differentiated 3T3-L1 adipocytes, which was associated with the inhibition of the expression of adipogenesis-related transcription factors, such as peroxisome proliferator-activated receptor γ, CCAAT/enhancer-binding protein-α (C/EBPα) , and C/EBPβ, which function as the key regulators of adipogenesis. Also, MECF reduced tyrosinase activity and melanin content in B16F10 cells as well as the expression of tyrosinase, tyrosinase-related protein-1 (TRP-1), TRP-2, and microphthalmia-related transcription factor in the presence of α-melanocyte-stimulating hormone. Taken together, our findings suggest that the extract of C. fragile could be considered a promising functional ingredient for the prevention and treatment of obesity and skin pigmentation in the food and cosmetic industry.

해양 미생물Algibacter lectus AS-3으로부터 agarase의 분리 및 특성

정일선,최영주,Jung, Il Sun,Choi, Young Ju 한국해양바이오학회 2007 한국해양바이오학회지 Vol.2 No.3

Agar로부터 oligosaccharides를 제조하기 위하여 효소 (agarase)를 이용한 분해법이 시도되면서 agarase의 균주의 분리 및 유전자에 대한 많은 연구가 수행되고 있다. 본 연구에서는 해양으로부터 agarase를 생성하는 균주를 분리 동정하고 균주의 특성을 조사하였다. 분리된 균주는 AS-3균주는 16S rDNA 염기서열분석에 의하여 Algibacter lectus AS-3로 동정되었다. 정제된 효소의 최적 활성 조건 및 agar 분해산물의 항산화활성을 조사하였다. 분리된 균주의 최적배양조건은 marine broth 2216에서 온도 $27^{\circ}C$, pH 7일 때 가장 균주의 생육이 높았다. Agarase 효소는 salt 침전, ion exchange와 gel filtration chromatography에 의해 9.6 units/mg으로 6.9배 정제되었다. 최적 효소활성은 온도 $40{\sim}50^{\circ}C$, pH 7일 때 나타났다. Agar 첨가한 배지에 agar 분해균을 접종한 후 분해산물의 시간에 따른 항산화활성은 12시간 배양 후 62%의 가장 높은 전자소거능 (EDA)을 나타내었다. An agar-degrading marine bacterium, strain AS-3 was isolated from the seawater. The strain AS-3 was identified as Algibacter lectus AS-3 by 16S rDNA sequence. The optimum medium for agarase activity of the isolated strain was determined to be marine medium, marine broth 2216 containing 0.1% agar as carbon source. An extracellular agarase was purified 6.9-fold from the culture supernatant by ammonium sulfate precipitation, ion exchange chromatography and gel filtration methods. The optimum pH and temperature for this enzyme were 7.0 and $40-50^{\circ}C$, respectively. Antioxidative activity of the strain AS-3 was 62.4% in the supernatant cultured for 12 h.

대황(Eisenia bicyclis) 배우체와 아포체의 생장에 미치는 환경 인자의 영향

이민정(Min-Jeong Lee),김남길(Nam-Gil Kim) 한국해양바이오학회 2020 한국해양바이오학회지 Vol.12 No.2

Eisenia bicyclis, a perennial macroalga is a primary producer of in the ocean, It has been identified as a key species that plays a vital role in maintaining the ecosystem stability. Also, it is an important target in marine afforestation projects and useful marine organisms. In addition, E. bicyclis is used as a health food for humans. This study investigated the effect of water temperature, light (photon irradiance), and duration of light (photoperiod) on the growth of gametophytes and young sporophytes of E. bicyclis. The germination and growth of the zoospores of E. bicyclis were examined at five temperatures (5℃, 10℃, 15℃, 20℃ and 25℃), four intensities of photon irradiance (10, 20, 40, and 80 μmol m<SUP>-2</SUP>s<SUP>-1</SUP>), and photoperiods (14:10 and 10:14 light/dark cycles). The zoospores released from mature plant germinated into the gametophytes under all experimental conditions. The gametophytes were able to grow at water temperature 5℃-25℃ and mature at 10℃-20℃. The optimal range of water temperature for the maturation of the gametophyte was 15℃-20℃. At 25℃, E. bicyclis gametophytes grew rapidly but did not mature. The optimal culture conditions for the growth of young sporophytes grew slowly in low temperature and photon irradiances.

미세조류 배양을 이용한 부영양호 내 수질 개선 기술 개발

김기현(Ki-Hyun Kim),강성모(Sung-Mo Kang),조용희(Yonghee Cho),전상현(Sanghyun Jeon),김준호(Jun-Ho Kim),박한울(Hanwool Park),이윤우(Yunwoo Lee),정정호(Jeongho Jeong),임상민(Sang-Min Lim),이철균(Choul-Gyun Lee) 한국해양바이오학회 2018 한국해양바이오학회지 Vol.10 No.2

There are many eutrophic lakes by point and non-point pollution sources such as industrial waste water, domestic raw sewage, and mucks. The eutrophic lakes not only cause algal blooms but also destroy the ecosystem in the lakes due to high nutrient concentrations. The purpose of this study was to improve water quality in eutrophic lakes by cultivating microalgae using photobioreactors (PBRs) with selectively permeable mesh (SPM), supplying nutrients in the lake and inhibiting cell leakage by diffusion and water permeability. Chlorella vulgaris, was cultivated using PBRs with SPM installed in Inkyung Lake located in Inha university, Incheon, Korea. When cultivating C. vulgaris, 8.3 g/㎡/day of average biomass productivity was obtained at 3 days. Furthermore, concentrations of total nitrogen and phosphorus were reduced by 35.7% and 84.2%, respectively, compared to initial condition and water quality in eutrophic lake was improved to oligotrophic environment. These results suggest that microalgal cultivation using PBRs with SPM in the lake could produce microalgal biomass as well as improve water quality by decreasing nutrient concentrations.

통연 연안에 분포하는 거머리말의 계절변동과 생식특성

윤준식,김남길 (사)한국해양바이오학회 2019 한국해양바이오학회지 Vol.11 No.2

We investigated the variation of morphological features, density, biomass and characteristics of reproductive shoot on Zostera marina. This species was monthly monitored and collected during a year in Tongyeong (from January 2016 to December 2016). Morphological features, biomass and density showed significantly seasonal variation (p<0.001). Dimensions of Z. marina were highest in spring and early summer but lowest in winter. Biomass and leaf density of Z. marina showed highest value in May (4,700.5 g w·wt m-2) and June (858.0 leaves m-2) respectively and lowest value in November (515.9 g w·wt m-2 and 312.0 leaves m-2). Reproductive shoot was monitored from April (13℃) to June (21℃) and grew up to ca. 200 cm. Spathe length, spadix weight, numbers and weight of fruit showed significantly different on monthly. None mature flowers were monitored in April but matured flowers, fruits were founded in May and seed releasing were occurred in June.

플라스크 배양에서 Chlorella sp. KR-1의 균체 성장 및 지질 생산에 대한 질소원 및 유기탄소원의 영향

이자연,서경애,오유관,Lee, Ja-Youn,Seo, Kyoung Ae,Oh, You-Kwan 한국해양바이오학회 2014 한국해양바이오학회지 Vol.6 No.2

Recently microalgae have been proposed as a promising biodiesel feedstock, owing to their higher lipid productivity and non-arable land based cultivation system. Biomass and lipid productivities of microalgae are largely affected by various environmental and nutritional factors. In this study, the effects of nitrogen (nitrate and ammonium) and organic carbon (glucose and glycerol) sources on the cell growth and lipid production of Chlorella sp. KR-1 were examined in flask cultures. Under autotrophic culture conditions for 15 days, overall cell growth and lipid (fatty acid methyl ester, FAME) production with nitrate were better than those of ammonium, resulting in 1.06 g cell/L and 333 mg FAME/L, respectively. Maximal intracellular lipid contents (348 - 352 mg FAME/g cell) were observed at low concentrations of 1 mM for both nitrate and ammonium. In the supply of light, addition of glucose in the range of 1 - 20 g/L showed higher cell densities than the autotrophic cell growth condition. Higher lipid accumulation of 375 mg FAME/g cell could achieved at 5 g glucose/L albeit of relatively short incubation of 7 days. With glycerol, intracellular lipid contents were ~1.9 times lower than glucose cases although similar cell growths were observed for both carbon sources.

Lentivirus System을 이용한 Glucocorticoid 유도 Reporter 유전자 발현의 분석

이미숙,김지연,허송욱,Lee, Mi-Sook,Kim, Ji-Yeon,Her, Song 한국해양바이오학회 2007 한국해양바이오학회지 Vol.2 No.2

글루코코르티코이드의 다양한 생리학적 과정은 이 호르몬에 의해 활성화된 수용체가 표적 유전자의 전사를 촉진 혹은 억제시킴으로써 일어나게 된다. 본 논문은 렌티바이러스 리포터 시스템을 이용하여 글루코코르티코이드 호르몬에 의한 GR 활성을 핵내에서 GRE에 의해 유도된 리포터 단백질인 mRFP 또는 루시퍼라아제의 발현을 통해 정성, 정량화 하였다. 그 결과 GR이 endogenous 하게 발현되는 HeLa 세포에서 코티졸을 처리하였을 때 활성화된 GR에 의해 GRE-inducible한 RFP와 루시퍼라아제의 발현이 각각 공초점 형광 현미경과 IVIS-200을 이용하여 형광 또는 BLI을 통해 증가함을 확인하였다. 이러한 결과를 통해 렌티바이러스 리포터 시스템을 이용한 연구는 세포 내에서 뿐 만 아니라 향후 생체내에서의 GR signaling을 모니터링하는데 유용하게 사용되어질 수 있을 것이다. Glucocorticoid hormone regulates numerous physiological processes, such as regulation of metabolism, and anti-inflammatory and immunosuppressive actions via the activation and repression of gene expression. Here we described a lentivirus-based reporter vector system expressing red fluorescent protein (mRFP) or firefly luciferase (Luc) under the control of a glucocorticoid-responsive element that allows observation of the temporospatial pattern of glucocorticoid induced GR-mediated signaling on a cellular level. Moreover, usage of the chromatin insulator of the chicken ${\beta}$-globin locus induced a marked increase of sensitivity of glucocorticoid inducible promoter of a reporter gene. Use of this method will be applicable of screening for agonist and antagonist of GR in vitro, and also a reporter gene assay for the in vivo determination of the GR-mediated gene activation.

황백의 주요 구성 화합물에 의한 약물대사효소 및 약물수송단백 저해능 평가

구혜영,김현미,손지홍,유광현,Ku, Hei-Young,Kim, Hyunmi,Shon, Ji-Hong,Liu, Kwang-Hyeon 한국해양바이오학회 2006 한국해양바이오학회지 Vol.1 No.3

본 연구는 황백에 함유되어 있는 주요 화합물인 berberine, palmatine, limonin 및 rutaecarpine의 CYP2D6 및 p-glycoprotein 활성에 대한 저해정도를 탐색함으로써, 황백을 다른 양약과 병용시 약물상호작용을 유발할 수 있는 가능성을 평가하고자 하였다. 인체 간 마이크로좀 시료에 CYP2D6 동효소의 기질약물인 dextromethorphan과 NADPH 재생성계 및 저해제 ($200{\mu}M$)를 첨가한 후 반응시켜 생성된 대사물을 LC/MS/MS를 이용하여 정량하여 CYP2D6 동효소 활성의 변화를 평가하였다. 또한 약물수송단백인 p-glycoprotein의 활성은 L-MDR1 세포주를 이용한 calcein AM 축적 실험을 통하여 평가하였다. 그 결과 식물 알카로이드인 berberine에서 강력한 CYP2D6 활성 저해능을 관찰하였으며, 저해 효과는 농도 의존적으로 증가하였으며, mechanism-based 저해 기전을 나타내었다. 그러나 limonine과 rutaecarpine은 CYP2D6 저해 활성을 보이지 않았고, p-glycoprotein 기능에 대해서는 평가한 어떤 화합물도 저해 활성을 나타내지 않았다. 황백의 주요 성분인 berberine의 CYP2D6 활성 저해능을 고려할 때, 황백을 CYP2D6 기질약제와 병용시 약물상호작용을 유발할 가능성을 보여준다. 이러한 황백의 CYP2D6를 매개로한 임상적인 약물상호작용 가능성은 임상시험을 통하여 추가적인 검정이 필요할 것으로 사료된다. We evaluated the potential of major components of Phellodendri cortex to inhibit the activities of CYP2D6 and p-glycoprotein. The abilities of berberine, palmatine, limonin, and rutaecarpine to inhibit CYP2D6-mediated dextromethorphan O-demethylation and calcein AM accumulation were tested using human liver microsomes and L-MDR1 cell, respectively. Berberine strongly inhibited CYP2D6 isoform activity, whereas limonin and reuaecarpine did not. The $IC_{50}$ value of berberine was reduced after preincubation with microsomes in the presence of NADPH generating system, suggesting that berberine is a mechanism based inhibitor. In addition, all chemicals tested, didn't show inhibitory effect on p-glycoprotein activity. These results suggest that berberine has potential to inhibit CYP2D6 activity in vitro. Therefore, in vivo studies investigating the interactions between berberine and CYP2D6 substrates are necessary to determine whether inhibition of CYP2D6 activity by berberine is clinically relevant.

대하 Penaeidin 3-2 유전자의 동정 및 발현

박은미,조현국,홍경은,남보혜,김영옥,김우진,이상준,한현섭,이재용,김종식,장인권,정재훈,최태진,공희정,Park, Eun-Mi,Cho, Hyun Kook,Hong, Gyeong-Eun,Nam, Bo-Hye,Kim, Young-Ok,Kim, Woo-Jin,Lee, Sang-Jun,Han, Hyon Sob,Lee, Jae Yong,Kim, Jong-Sheek,Ja 한국해양바이오학회 2007 한국해양바이오학회지 Vol.2 No.1

Penaeidins은 새우류에 존재하는 항미생물성펩티드의 한 종류로서 새우의 외부 병원체에 대한 방어기작을 구성하는 중요한 인자이다. 본 연구에서는 대하 혈구세포의 cDNA library로부터 분리된 EST 클론을 분리 동정하였다. 아미노산 염기서열 분석과 계통수 분석을 통하여 본 연구에서 분리한 EST 클론이 대하의 Penaeidin 3-2 유전자와 아미노산 수준에서 동일함을 밝혔다. 대하의 Penaeidin 3-2는 signal peptides로 예상되는 19개 아미노산 잔기를 포함하여 전체 71개의 아미노산 잔기로 이루어져 있고, C-말단에는 3개의 이중황화결합을 형성할 수 있는 6개의 cysteine가 존재하였다. 대하 Penaeidin 3-2 전사체는 혈구세포, 간췌장, 근육 조직에서 발현되었으며, 특히 혈구세포에서 주로 많이 발현되었다. 흰반점바이러스의 인위감염 실험에서 바이러스 감염 후 대하 Penaeidin 3-2의 발현이 증가하였다. 이상의 결과들로부터 대하 Penaeidin 3-2가 자체 방어 작용에서 중요한 역할을 할 것으로 예상된다. Penaeidins are members of a special family of antimicrobial peptides existing in several species of shrimp and play a crucial role in the immunological defense of shrimp. In this study, we isolated and characterized one EST clone (penaeidin) from cDNA library of fleshy prawn Fenneropenaeus chinensis hemocytes. Amino acids sequence comparison and phylogenetic analysis with other known penaeidins revealed that our clone was completely identical to F. chinensis Penaeidin 3-2 (Accession no. ABC33920), which composed of 71 amino acids with a putative signal peptide (1-19) and a cysteine-rich domain (C-terminal part). The expression and distribution of Penaeidin 3-2 transcripts in shrimp were detected in hemocytes, hepatopancreas, and muscles, and that Penaeidin 3-2 was constitutively expressed mainly in hemocytes. The artificial infection of white spot syndrome virus to F. chinensis resulted in Penaeidin 3-2 mRNA up-regulation in hemocytes, suggesting that the possible role of Penaeidin 3-2 in host defense system of F. chinensis.