• KSBB Journal
• /
• 제20권3호
• /
• pp.233-237
• /
• 2005

누에체액 성분 중의 30K 단백질은 Q-Sepharose FF를 이용한 단계별 농도구배 이온교환 크로마토그래피로 0.16 M NaCl 용출 조건에서 쉽게 얻을 수 있었다. 얻어진 단백질을 인간유래 정상세포에 처리하여 세포 증식 향상이 확인되었고, 이는 30K 단백질이 인간을 위한 세포치료제에 이용되는 세포의 증식을 위한 첨가물로 사용될 수 있음으로써 의료 분야에의 적용 가능성을 확인하였다. 또한 본 연구를 통해 확립된 30K 단백질의 간단한 정제 공정은 향후 대량 생산 공정으로 scale-up이 용이하게 이루어질 것으로 기대된다. 이 결과들은 동물세포를 생산 숙주로 사용할 때 생산 물질 자체의 생산량 증대에도 효과적으로 이용될 수 있으며 세포치료제를 위한 세포의 증식에도 효과를 보일 수 있으므로 세포배양을 이용한 생산 공정 전반에 큰 영향을 미칠 수 있을 것이다.

• 한국균학회소식:학술대회논문집
• /
• 한국균학회 2015년도 추계학술대회 및 정기총회
• /
• pp.41-41
• /
• 2015

Oomycetes belong to the kingdom Straminipila, a remarkably diverse group which includes brown algae and planktonic diatoms, although they have previously been classified under the kingdom Fungi. These organisms have evolved both saprophytic and pathogenic lifestyles, and more than 60% of the known species are pathogens on plants, the majority of which are classified into the order Peronosporales (includes downy mildews, Phytophthora, and Pythium). Recent phylogenetic investigations based on DNA sequences have revealed that the diversity of oomycetes has been largely underestimated. Although morphology is the most valuable criterion for their identification and diversity, morphological species identification is time-consuming and in some groups very difficult, especially for non-taxonomists. DNA barcoding is a fast and reliable tool for identification of species, enabling us to unravel the diversity and distribution of oomycetes. Accurate species determination of plant pathogens is a prerequisite for their control and quarantine, and further for assessing their potential threat to crops. The mitochondrial cox2 gene has been widely used for identification, taxonomy and phylogeny of various oomycete groups. However, recently the cox1 gene was proposed as a DNA barcode marker instead, together with ITS rDNA. To determine which out of cox1 or cox2 is best suited as universal oomycete barcode, we compared these two genes in terms of (1) PCR efficiency for 31 representative genera, as well as for historic herbarium specimens, and (2) in terms of sequence polymorphism, intra- and interspecific divergence. The primer sets for cox2 successfully amplified all oomycete genera tested, while cox1 failed to amplify three genera. In addition, cox2 exhibited higher PCR efficiency for historic herbarium specimens, providing easier access to barcoding type material. In addition, cox2 yielded higher species identification success, with higher interspecific and lower intraspecific divergences than cox1. Therefore, cox2 is suggested as a partner DNA barcode along with ITS rDNA instead of cox1. Including the two barcoding markers, ITS rDNA and cox2 mtDNA, the multi-locus phylogenetic analyses were performed to resolve two complex clades, Bremia lactucae (lettuce downy mildew) and Peronospora effuse (spinach downy mildew) at the species level and to infer evolutionary relationships within them. The approaches discriminated all currently accepted species and revealed several previously unrecognized lineages, which are specific to a host genus or species. The sequence polymorphisms were useful to develop a real-time quantitative PCR (qPCR) assay for detection of airborne inoculum of B. lactucae and P. effusa. Specificity tests revealed that the qPCR assay is specific for detection of each species. This assay is sensitive, enabling detection of very low levels of inoculum that may be present in the field. Early detection of the pathogen, coupled with knowledge of other factors that favor downy mildew outbreaks, may enable disease forecasting for judicious timing of fungicide applications.

• 한국식품영양과학회지
• /
• 제17권3호
• /
• pp.226-232
• /
• 1988

변온조건하에서 녹차를 저장하였을 때 수분활성에 따른 browning development를 반응속도론적으로 고찰한 결과 brownung development는 영차반응으로 증가하였으며, 반응속도는 수분활성이 높을수록, 저장속도가 높을수록 빨랐고, 각 수분활성에서의 활성화 에너지는 $1.5{\sim}2.4kcal/mole$, $Q_{10}$치는 $1.07{\sim}1.12$였다. Accelerated shelf-life test로부터 구한 $25^{\circ}C$에서의 shelf-life는 $57{\sim}113$일의 범위였으며, 온도와 수분활성이 증가함에 따라 단축되었다. 변온조건에서의 실측치와 예측치를 비교한 결과 유효온도차는 $2.66{\sim}5.64^{\circ}C$였고, shelf-life는 예측치가 높게 나타났으나 이 방면의 연구가 더욱 진행된다면 변온저장의 결과를 효율적으로 예측할 수 있을 것으로 예상된다.

• 식물병연구
• /
• 제18권3호
• /
• pp.201-209
• /
• 2012

지자체 농업기술센터와 농과원에서 수집한 18개의 Bacillus subtilis 균주들이 보유하고 있는 기능성을 구명하기 위하여 항균활성을 비롯한 사이데로포아 생산, 질소고정능력, 인산가용화 능력 및 오옥신 생성능력을 조사하였다. B. subtilis 균주들은 대부분 입고병균(R. solani), 고추 탄저병균(C. acutatum), 채소류 시들음병균(F. oxysporum), 벼도열병균(M. oryzae), 고추 역병균(P. capsici)에 대하여 항균활성을 보였다. 대부분의 균주가 Siderophore와 오옥신 생성능력을 보였으며 질소고정능력도 함께 보유하고 있는 것으로 나타났다. 인산가용화능은 조사한 Bacillus 속 균주 중 단 한 균주에서 활성을 보였다. 대부분의 B. subtilis 균주가 고추 흰가루병 억제효과를 보였으나, 균주 간에는 방제효과에 있어서 큰 차이를 보이지 않았다. B. subtilis R2-1균주의 농업현장에서의 활용 가능성을 구명하기 위하여 2009년도에 강원도 화천의 유기재배농가에서 오이 흰가루병 방제용 미생물 농약으로 등록되어 사용되고 있는 '큐펙트'와 '탑시드'를 대조로 하여 방제효과를 검정하였다. 2009년도에 B. subtilis R2-1 처리시 방제효과는 37.7%로 '탑시드'(47.6%)보다는 낮았으나, '큐펙트'(25.7%)에 비해서는 높았다. 2010년도에는 B. subtilis R2-1의 고추 흰가루병 방제효과가 83.3%로 미생물 농약인 탑시드 보다도 높게 나타났다. B. subtilis R2-1균주의 발병억제 기작을 구명하기 위하여 B. subtilis R2-1균주와 미생물 농약(탑시드, 큐펙트)을 경엽 처리한 후 포자발아율을 계수한 결과, 미생물 농약인 큐펙트는 97.9%, 탑시드는 94.7% 억제하는데 비해서 B. subtilis R2-1균주는 84.2% 억제하는 것으로 나타났다. 앞으로 B. subtilis R2-1을 고농도로 배양할 수 있는 대량배양기술을 개발하고 항균활성 기능이 잘 발현되도록 제형화한다면 생물농약으로의 개발이 가능할 것으로 사료된다.

• Applied Biological Chemistry
• /
• 제20권1호
• /
• pp.66-80
• /
• 1977

사과 증류주(apple One spirit) 숙성에 있어서 국산 참나무의 품종이 fine spirit숙성 술통재로서의 적성여부를 검토하는 기초적 연구의 일환으로 본 연구를 실행하여 다음과 같은 결과를 얻었다. 1. 양조 원료를 대체 할 수 있는 과실로서 현재 대량생산 되는 과실의 화학분석 결과는 다음과 같다. 사과 : (Apple) (Malus pumila Miller var domestica Schneider) 중 홍옥(Jonathan)은 총당 13.95%, 총산 0.46%, 휘발산 0.012%, pectin 0.20%, 국광(Ralls)은 총당 13.35%, 총산 0.43%, 휘발산 0.011%, pectin 0.455%이었다. 2. 사과중에는 당분과 산 이외에 cellulose, pectin, hemicellulose 등 때문에 사과즙수을이 떨어짐으로 Aspergillus niger SUAFM-430의 xylanase와 amylase, Aspergillus niger SUAFM-6의 cellulase, pectinase를 사과즙에 처리하여 사과즙의 수율을 향상시켰다. 그리고 이황산 처리로 과즙중 여러 미생물을 살균하고 배양효모로서 발효능이 우수한 sacharomyces cerevisiae var. ellipsoidens Rasse Johannisberg II (SUAFM-1018)을 사용하여 사과주를 양조하였다. 사과주(Apple wine)는 원료에 대한 수율이 $86{\sim}87%$이고 Jonathan, Ralls가 ethanol 13.5%, extract분 5.4%, methanol $0.04{\sim}0.05%$이었다. 3. 사과주의 증류수(fine spirit) 제조는 단식증류법(Pot still)으로 2회 증류로서 실험하였다. 사과(apple) fine spirit는 apple wine mash에 대한 수율이 86.6%이고, 홍옥(Jonathan) fine spirit는 pH 4.1, 국광(Ralls) fine spirit는 pH 4.2이었다. 4. 사과주를 증류하여 제조한 fine spirit를 숙성시키기 위하여 국산 참나무 24품종을 나무의 심재(心材) (Inner part)와 변재(邊材) (Outer part)를 사용하여 fine spirit의 숙성융통제를 선발하는 실험을 하였다. 참나무 24품종과 그의 심재, 변재 별로 참나무의 절편(oaa chip) $(1{\times}1{\times}5cm)$을 만들고 oakchip 2개를 각 fine spirit 300ml씩 들은 640ml용맥주병에 담그고, 6개월간 숙성시키면서 색, 향기, 맛 등을 조사하였다. A. 숙성용술통재 선발의 1차 screen결과로서 색추출속도가 우수한 참나무 품종은 다음 순서와 같다. 갈참나무(심재) Quercus aliena Blume(Inner part), 굴참나무(변재) Q variabilis Blume (Outer part), 갈참나무(변재) Q. aliena Blume (Outer part), 졸참나무(심, 변재) Q. serrata Thumb (Inner & Outer part), 신갈참나무(변, 심재) Q. mongolca Fisher (Outer & Inner part), 상수리 나무(변, 심재) Q. acutissima Carruthers (Outer & Inner part)의 순서등 5품종을 선정하였다. B. 숙성온도별 영향으로서 실온 $24{\sim}25^{\circ}C,\;30^{\circ}C,\;45^{\circ}C$ 별로 각 fine spirit에 oak chip을 넣고 숙성시킨 결과 $45^{\circ}C$에서 숙성시킨 것이 가장 숙성이 촉진 되었고 다음이 $30^{\circ}C$, 다음이 실온에서 한 순서의 경향이었다. C. 투명한 삼각후라스크에 각 fine spirit에 oak chip을 담그고 자외선 조사를 하여 3개월간 숙성 시켰드니 숙성효과가 contral구 보다 거의 2배가 촉진하는 흥미있는 사실을 확인하였다. D. 사과주에서 증류한 fine spirit 300ml에 2개의 oak chip을 담그어 숙성시킴에 있어 oak chip의 여러성분이 fine spirit에 용출되여 나오는 정도가 fine spirit의 pH에 중요한 관계가 있음을 알았다. E. 선정된 5개 품종의 oak chip과 블란서 참나무 술통재(Limousin white oak from France)를 control로 하여 각 사과주를 증류한 fine spirit에 2개의 oak chip($1{\times}1{\times}5cm$)을 담그고 실온, $(24{\sim}25^{\circ}C)$, $30^{\circ}C,\;45^{\circ}C$, 실온에서 자외선조사 하면서 각각 6개월간 매일주일 마다 진탕하면서 숙성시켰다. 6개월간 수성시킨 fine spirit(young brandy)의 관능 검사 결과는 다음과 같다. 불란서 oak chip의 실온 숙성한 fine spirit 하고 비슷한 것은 참갈나무 chip을 넣은 사과의 young bandy($45^{\circ}C$에서 숙성함)이었고 갈참나무 chip을 넣고 실온에서 UV-ray 조사한 사과의 young brandy는 불란서 oak chip의 실온 숙성한 young brandy 보다는 약간 떨어지지만 그런대로 국산 참나무를 각 fine spirit 숙성용으로 이용할 가치가 있음을 알았다.

• 한국환경과학회지
• /
• 제24권12호
• /
• pp.1591-1600
• /
• 2015

The response of the freshwater microalga, Chlorella vulgaris, to heavy metal stress was examined based on chlorophyll fluorescence analysis to assess the toxic effects of heavy metals in freshwater ecosystems. When toxic effects were analyzed using regular chlorophyll fluorescence analysis, photosystem II activity($F_v/F_m$) decreased significantly when exposed to $Cu^{2+}$ and $Hg^{2+}$ for 12 h, and decreased in the order of $Hg^{2+}>Cu^{2+}>Cd^{2+}>Ni^{2+}$ when exposed for 24h. The effective photochemical quantum yield(${\phi}{\prime}_{PSII}$), chlorophyll fluorescence decrease ratio($R_{Fd}$), minimal fluorescence yield($F_o$), and non-photochemical quenching(NPQ), but not photochemical quenching(qP), responded sensitively to $Hg^{2+}$, $Cu^{2+}$, and $Cd^{2+}$. These results suggest that $F_v/F_m$, as well as ${\phi}{\prime}_{PSII}$, $R_{Fd}$, $F_o$, and NPQ could be used to assess the effects of heavy metal ions in freshwater ecosystems. However, because many types of heavy metal ions and toxic compounds co-occur under natural conditions, it is difficult to assess heavy metal toxicity in freshwater ecosystems. When Chlorella was exposed to heavy metal ions for 12 or 24h, $F_v/F_m$ and maximal fluorescence yield($F_m$) changed in response to $Hg^{2+}$ and $Cu^{2+}$ based on image analysis. However, assessing quantitatively the toxic effects of several heavy metal ions is challenging.

• 한국발생생물학회지:발생과생식
• /
• 제24권1호
• /
• pp.43-52
• /
• 2020

NUCB2/nesfatin-1 known to regulate appetite and energy homeostasis is expressed not only in the hypothalamus, but also in various organs and tissues. Our previous reports also demonstrated that NUCB2/nesfatin-1 was expressed in the reproductive organs, including the ovaries, uterus, and testes of mice. However, it is yet known whether NUCB2/nesfatin-1 is expressed in the oviduct and how its expression is regulated. Therefore, we investigated the expression of NUCB2/nesfatin-1 in the oviduct and its expression is regulated by gonadotropin. Immunohistochemical staining results showed that nesfatin-1 protein was localized in epithelial cells of the oviduct. As a result of quantitative real-time PCR (qRT-PCR) and Western blot, NUCB2/nesfatin-1 was detected strongly in the oviducts. During the estrus cycle, NUCB2/nesfatin-1 expression in the oviducts was markedly higher in the proestrus stage than in other estrus stages. In order to elucidate whether the expression of NUCB2 mRNA is controlled by the gonadotropins, we injected PMSG and hCG and measured NUCB2 mRNA level in the oviduct after injection. Its level was increased in the oviduct after PMSG injection, but no significant change after hCG injection. In addition, NUCB2 mRNA levels were markedly reduced after ovariectomy, while recovered after 17β-estradiol (E2) injection, but not by progesterone (P4). This study demonstrated that NUCB2/nesfatin-1 is highly expressed in the oviduct of mouse and its expression is regulated by E2 secreted by the ovaries. These results suggest that NUCB2/nesfatin-1 expressed by the oviduct may affect the function of the oviduct regulated by the ovaries.

• Journal of Microbiology and Biotechnology
• /
• 제18권6호
• /
• pp.1076-1080
• /
• 2008

Glucagon, a peptide hormone produced by alpha-cells of Langerhans islets, is a physiological antagonist of insulin and stimulator of its secretion. In order to improve its bioactivity, we modified its structure at the C-terminus by amidation catalyzed by a recombinant amidase in bacterial cells. The human gene coding for glucagon-gly was PCR amplified using three overlapping primers and cloned together with a rat ${\alpha}$-amidase gene in plasmid pMGA. Both genes were expressed under control of the strong constitutive promoter of aph and secretion signal melC1 in Streptomyces lividans. With Phenyl-Sepharose 6 FF, Q-Sepharose FF, SP-Sepharose FF chromatographies and HPLC, the peptide was purified to about 93.4% purity. The molecular mass of the peptide is 3.494 kDa as analyzed by MALDI TOF, which agrees with the theoretical mass value of the C-terminal amidated glucagon. The N-terminal sequence of the peptide was also determined, confirming its identity with human glucagon at the N-terminal part. ELISA showed that the purified peptide amide is bioactive in reacting with glucagon antibodies.

• 한국전기전자재료학회논문지
• /
• 제17권12호
• /
• pp.1320-1325
• /
• 2004

In this study, in order to develop the low temperature sintering ceramics for multilayer piezoelectric transformer, PCW-PMN-PZT ceramics added with Li$_2$CO$_3$, Bi$_2$O$_3$ and CuO as sintering aids were manufactured, and their microstructural, dielectric and piezoelectric properties were investigated. When the only CuO was added, specimens could not be sintered below 98$0^{\circ}C$. However, when Li$_2$CO$_3$ and Bi$_2$O$_3$ were added, specimens could be sintered below 98$0^{\circ}C$. Li$_2$CO$_3$ and Bi$_2$O$_3$ addition were proved to lower sintering temperature of piezoelectric ceramics due to the effect of Li$_2$O-Bi$_2$O$_3$ liquid phase. Li$_2$CO$_3$ and Bi$_2$O$_3$ added specimens showed higher piezoelectric properties than those of the only CuO added specimens. At 0.2 wt% Li$_2$CO$_3$ and 0.3 wt% Bi$_2$O$_3$ added specimen sintered at 92$0^{\circ}C$, the dielectric constant of 1457, electromechanical coupling factor of 0.56 and mechanical quality factor of 1000 were shown, respectively. These values are suitable for multilayer piezoelectric transformer application.

• Molecules and Cells
• /
• 제41권3호
• /
• pp.179-187
• /
• 2018

Proteomic analysis of extracellular vesicles (EVs) from biological fluid is a powerful approach to discover potential biomarkers for human diseases including cancers, as EV secreted to biological fluids are originated from the affected tissue. In order to investigate significant molecules related to the pathogenesis of bladder cancer, EVs were isolated from patient urine which was analyzed by mass spectrometry based proteomics. Comparison of the EV proteome to the whole urine proteome demonstrated an increased number of protein identification in EV. Comparative MS analyses of urinary EV from control subjects and bladder cancer patients identified a total of 1,222 proteins. Statistical analyses provided 56 proteins significantly increased in bladder cancer urine, including proteins for which expression levels varied by cancer stage (P-value < 0.05). While urine represents a valuable, non-invasive specimen for biomarker discovery in urologic cancers, there is a high degree of intra- and inter-individual variability in urine samples. The enrichment of urinary EV demonstrated its capability and applicability of providing a focused identification of biologically relevant proteins in urological diseases.