• 한국식품과학회지
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• 제29권1호
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• pp.145-149
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• 1997

쌀의 저장시 지방질의 가수분해나 산화에 의해 off flavor를 생성하여 쌀의 품질을 저하시키는데 있어 촉매작용을 하는 lipoxygenase 활성을 비색법으로 측정하였다. 기질농도와 효소반응속도와의 상호관계를 검토하고, 최적 pH를 조사하였으며, 도정도에 따른 현미 각 부분의 효소활성을 알아보았다. 0.2 mM의 linoleic acid/phosphate buffer (pH 7.0)와 반응시켰을 때 반응시간에 따른 효소활성은 동진이 3시간, 금오가 4시간, 간척이 5시간까지 효소활성이 증가하였다. 기질의 농도와 효소액과의 반응속도 관계를 Lineweaver-Burk plotting으로 해석한 결과 $V_{max}$는 동진, 금오 및 간척벼는 각각 57.89, 19.85, 31.38 unit/mg protein이었고, $K_m$은 동진, 금오 및 간척벼 각각 0.054, 0.045, 0.035 mM이었다. 현미의 lipoxygenase의 활성은 동진벼의 경우 pH 7.6, 간척벼는 pH 7.0, 금오벼는 pH 7.0에서 최적의 활성을 나타내었다. 도정도에 따른 현미 각 부분의 효소활성 및 비활성은 동진, 금오 및 간척벼 모두 획분 II에서 가장 높은 활성을 보였으며 획분 IV에서 가장 낮은 값의 활성을 보였다. 세 품종의 현미에 microwave로 0, 30, 60, 90초간 가열처리하였을 때 효소활성 및 효소의 비활성은 대조구에 비해 90초간 가열처리했을 때 효소활성의 경우 동진, 금오 및 간척벼는 27.4%, 52.2%, 17.1%로 감소하였으며, 효소의 비활성도 42.6%, 76.7%, 23.4% 정도로 감소하였다.

• 한국식품과학회지
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• 제6권2호
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• pp.75-78
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• 1974

본(本) 실험(實驗)에서는 현재(現在)까지 학문적(學問的)으로 충분(充分)히 밝혀지고 있지 않은, 한우유(韓牛乳)의 이화학적(理化學的) 성질(性質)을 구명(究明)하기 위(爲)하여 한우유(韓牛乳)의 주성분(主成分)을 분석(分析)하고, 또한 DEAE-cellulose를 사용(使用)하여 산(酸) casein, 유청단백질(乳淸蛋白質)을 분별(分別)하여 다음과 같은 결과(結果)를 얻었다. 1. 한우유(韓牛乳)의 비중(比重)은 1.036, 산도(酸度) 0.21, pH는 6.4였고 알콜시험(試驗)에서는 정상(正常)이었다. 2. 한우유(韓牛乳)의 수분함량(水分含量)은 88.39%였고, 무지고형분(無脂固形分) 9.53%였고, 지방(脂肪)은 2.08%, 단백질(蛋白質)은 3.99%, 유당(乳糖)은 4.76%였고 Sialic acid는 0.25%였다. 3. 총단백질중(總蛋白質中) casein이 3.07% 유청단백질(乳淸蛋白質)이 1.13%, lactoglobulin 0.34%, lactalbumin은 0.66%였다. 4. 6M 요소(尿素)를 함유(含有)한 0.005M TCU-buffer를 사용(使用)하여 DEAE-cellulose column에 의(依)해서 한우유(韓牛乳)의 산(酸) casein을 분별(分別)한 결과(結果) 4개(個)의 fraction을 얻었다. 5. 0.04M phosphate-buffer를 사용(使用)하여 DEAE-cellulose column에서 유청단백질(乳淸蛋白質)을 분별(分別)한 결과(結果) 4개(個)의 fraction을 얻어 각(各) fraction의 면적(面積)을 냈다.

• 약학회지
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• 제55권3호
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• pp.227-233
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• 2011

We previously reported the protein isolated from Coptidis Rhizoma (CRP), which has antifungal activity against a fungal pathogen, Candida albicans. In the current study, we investigated what portion in the CRP is responsible for the antifungal activity. For the investigation, the CRP was fractionated on a Shepadex G-50 column. Data resulting from the fractionation, seven fractions were obtained. Fractions (Fr.) I, II, and III eluted initially from the column showed no inhibitory effect on the growth of C. albicans, whereas Fr. IV, V, and VI eluted later revealed inhibition of the growth, and Fr. IV and VI showed potent antifungal activity by broth susceptibility analysis. However, Fr. VI was contained in the CRP more than Fr. IV, which led us to select the VI for the following experiments. In a murine model of a subcutaneous candidiasis caused by C. albicans, the Fr. VI displayed a therapeutic effect on nude mice pretreated with anti-neutrophil monoclonal antibody (RB68C5) and then infected subcutaneously with live C. albicans. At day 16, these mice were healed almost up to 78% of the infected area when compared to infected area of control nude mice that received diluent (Dulbecco's Phosphate-Buffered Saline; DPBS), instead of the Fr. VI (P<0.01). The Fr. VI blocked hyphal formation from blastoconidial form of C. albicans (P<0.01), which might prevent penetration of hyphae to the deeper site of skin and thus helps the healing. In the ionic strength test, the effect of Fr. was influenced by $Ca^{2+}$ ion just like other known antimicrobial peptides, but the influence was affected at an extremely high concentration such as 500 mM. Thus, such ion-concentration is considered to be meaningless in the clinical situation. Considering all data together, Coptidis Rhizoma is appeared to produce an antimicrobial peptide that has therapeutic effect on subcutaneous infection caused by C. albicans.

• Parasites, Hosts and Diseases
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• 제21권1호
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• pp.49-57
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• 1983

The distribution and Properties of branched chain amino acid aminotransferase (EC 2.6. 1.42) was investigated in adult Fasciola hepatica. Fascicla hepatica was fractionated by differential centrifugation into nuclear, mitochondrial and cytosolic fractions. The activity of branched chain amino acid aminotransferase was measured by the method of Ichihara and Koyama (1966) . Isozyme patterns of this enzyule was also examined by DEAE-cellulose column chromatography. The results obtained were as follows; 1. The activity in homogenate was found to be 12.69 units/g wet tissue. The activity of this enzyme was relatively high compared with those in rat tissues. 2. The distribution of branched chain amino acid aminotransferase in the subcellular organelles showed that 87.8% of the activity was in cytosolic, 10.9% in mitochondrial and 1.3% was in nuclear fraction. 3. Cytosolic fraction of Fasciola hepatica contained Enzyme I, but not Enzyme II and III, of branched chain amino acid aminotransferase. Ensyme I was eluted by 50mM phosphate buffier from DEAE-cellulose column and catalyzed the transamination of all three branched chain amino acids. 4. The Enzyme I was purified about 22-folds increase in specific activity after chromatography on DEAE-cellulose. 5. The best substrate among three amino acids (leucine, isoleucine and valise) was L-isoleucine. 6. The optimal temperature of Enzyme I was $45^{\circ}C$ and the optimal pH was 8.2. 7. The Km value for leucine of Enzyme I was 4.17 mM. 8. The Km values for a-ketoglutarate and pyridoxal phosphate of Enzyme I were 0.41mM and $4.76{\times}10^{-3}{\;}mM$, respectively.

• 생명과학회지
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• 제19권8호
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• pp.1062-1066
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• 2009

N-Acetylglucosamine kinase (GlcNAc kinase or NAGK; EC 2.7.1.59)는 GlcNAc를 인산화하여 GlcNAc-6-phosphate를 만드는 효소이다. 효소자체에 대한 자세한 연구에도 불구하고 포유류에서NAGK의 표현에 대한 연구는 거의 없다. 배양한 흰쥐의 해마신경세포에서 NAGK은 세포체/가지돌기 영역에서 클러스터(cluster)를 형성한다. 본 연구에서는 가지돌기의 긴 축에서부터 가쪽으로 돌출되는 NAGK 클러스터에 대하여 연구하였다. 배양한 해마신경세포를 NAGK와 다양한 연접표지단백질에 대한 항체로 이중염색한 결과 NAGK 클러스터가 가지돌기의 바닥에는 있었지만 억제성 연접후부위에는 존재하지 않았다. 또한 흰쥐 전뇌(forebrain)의 균질액(homogenate, BH), 연접체(synaptosome, S), 연접후치밀질(postsynaptic density, PSD) 분획을 NAGK 항체로 면역염색한 결과 NAGK는 연접체에는 있었지만 PSD 분획에는 존재하지 않았다. 이러한 결과들은 NAGK가 가지돌기가시(spine)의 바닥쪽으로 돌출됨을 의미한다.

• 한국식품저장유통학회지
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• 제11권3호
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• pp.417-423
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• 2004

BAAG의 첨가배지상에서 미생물세포의 생육이 크게 억제된다는 실험결과를 토대로, 미생물에 대한 항균작용의 방식을 알아 보고자, Botrytis cinerea를 배양하여 분리한 여러 가지 효소 활성에 미치는 영향을 조사하였다. 특히 에너지 대사와 관련된 효소들에 대한 영향을 살펴보기 위하여 여러가지 대사경로에서 주요한 역할을 하는 효소인, hexokinase,glucose 6-phosphate dehydrogenase, malate dehydrogenase와succinate dehydrogenase에 대한 영향을 살펴 본 결과, BAAG가 외ucose-6-phosphate dehydrogenase 및 malate dehydrogenase의활성에는 별다른 효과를 나타내지 않았으나 succinate dehydrogenase 및 hexokinase 효소활성에서는 약간의 효소활성 억제작용을 관찰할 수 있었다. 또한, 세포막에 미치는 영향을 알아 보기 위하여 $\beta$-galactosidase의 인위적 기질인 ONPG의 분해 여부를 조사한 결과, BAAG는 세포막의 기능에 영향을 끼침을 관찰하였다. BAAG의 항균 성분을 분리확인 하기 위해서 silica gel column chromatography를 하여 ethyl ether분획물에서 3개, ethyl acetate분회물에서 4개의peak를 얻었으며, 각 용매분획물에서 1개씩의 분획에서 강한항균 활성을 검출하였다. 이 분획을 NMR 및 fast atomicbombardment측정기에 의해서 분석한 결과, 화학구조를limonin과 naringin으로 동정할 수 있었다. 이상의 연구결과로 미루어, 본 연구에서 그 기능과 작용을 구명한 천연항균소재 BAAG는 수확한 과채류의 직접 처리제재 및 항균 포장 소재로 과채류의 선도유지를 목적으로 활용하여 그 효과를 기대할 수 있어, 과채류의 생산기반 보호와 농가소득에큰 도움이 될 것으로 생각된다.

• 한국세라믹학회지
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• 제55권6호
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• pp.590-596
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• 2018

To improve the injection behavior of brushite cement, dense ${\beta}-Ca_3(PO_4)_2$ (${\beta}-TCP$) granules were added to the starting material. The spherical ${\beta}-TCP$ granules prepared by spray-drying and subsequent sintering at $1000{\sim}1200^{\circ}C$ accounted for fractions of from 0.5 to 0.7 of the total ${\beta}-TCP$. The injection behavior was evaluated by measuring the injected mass divided by the loaded mass of paste in the syringe pump. The injected amount was increased with the increase in the fraction and sintering temperature of ${\beta}-TCP$ granules, except at $1200^{\circ}C$. The increase in the fraction of ${\beta}-TCP$ and its sintering temperature resulted in a decrease in the plastic limit, which is the volume of water required to liquefy the compact. The rest water could be utilized in the cement with the reduced plastic limit for improved injectability. The amounts of rest water assigned for powdery phase were estimated, and correlated with the injectability of paste.

• Journal of Microbiology and Biotechnology
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• 제15권4호
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• pp.767-771
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• 2005

Expression in yeast may prove more amenable to generating large amounts of viral antigens for a vaccine candidate. We, therefore, cloned the gene encoding the Hepatitis C virus (HCV) structural proteins (C-El-E2, c740) fused in-frame with, and immediately 3' to, the chicken-lysozyme signal peptide (C-SIG) gene and under the control of the yeast glyceraldehyde-3-phosphate dehydrogenase gene promoter. In yeast, the HCV structural proteins were expressed in two different forms: a processed and a nonprocessed aggregated form. Biophysical characterization by sucrose linear gradient centrifugation revealed that both forms were present in the same fractions with a buoyant density of 1.127-1.176 g/$cm^3$. These findings suggest that the efficient synthesis of HCV structural proteins in yeast may be an important tool to study virus assembly and may lead to the development of an HCV vaccine.

• 한국미생물·생명공학회지
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• 제20권6호
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• pp.699-703
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• 1992

본 연구에서는 간염치료제로 효과가 있는 민간제제(NS-P)의 성분 분석 및 항균 활성과 항산화 활성을 연구하였다. 이 NP-S는 그람 양성균인 Staphylococcus aureus와 그람 음성균인 Salmonella typhimurium 그리고 진균인 Candida albcans에 항균 활성을 보여 주었을 뿐만 아니라, 지질의 과산화를 억제하는 항산화 활성을 나타냈다. 이러한 NP-S는 C/H/N/S의 원소비가 15.5/4.8/11/10으로 황(S)이 상당히 많이 존재하고 있으며, 휘발성 물질인 암모니아를 2.11M 함유하고 있었다. 이 외에도 6.7의 펩타이드, 0.3의 유리 아미노산, 0.1의 무기 인산염을 함유하고 있으며, 미량 원소로서 Cl, Si, Mg, Zn도 함유하고 있다.

• Journal of Pharmaceutical Investigation
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• 제32권4호
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• pp.285-290
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• 2002

In order to assay the efficacy of newly synthesized antiviral compounds, in vitro studies of their active intracellular phosphorylated metabolites were established as compared with Zidovudine (ZDV). Antiviral base analogs require intracellular phosphorylation prior to the inhibition of HIV replication. Therefore, antiviral drugs concentrations in plasma have not reflected any direct relationship with activity or toxicity. A method has been developed to measure the concentration of total phosphorylated metabolites inside peripheral blood mononuclear cells using modified commercial radioimmunoassay (RIA). ZDV 5'-monophosphate was synthesized and used as a procedural control for RIA modification. PBMCs were isolated from whole blood and incubated with ZDV for 20 h to allow metabolic phosphorylation. Viable cells were extracted overnight with 60% methanol. After evaporation, the extract was reconstituted in Tris buffer. Samples were split into two fractions, one of which was treated with alkaline phosphatase (AP) to liberate phosphate groups. Concentrations of phosphorylated metabolites were determined by subtracting thε concentration of non-AP-treated fraction from that of the treated fraction. Recovery of phosphorylated ZDV from cell extracts was approximately 90%, and reproducibility was acceptable (coefficients of variation <15% for concentrations${\geq}$0.25 ng/mL). Intracellular concentrations $(0.135{\sim}5.019\;nmole/10^6\;cells)$ followed a nonlinear dose-response relationship over the range $0.015{\sim}2.996mM$ extracellular ZDV, with concentration-dependant saturation.