• Applied Biological Chemistry
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• 제17권2호
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• pp.138-142
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• 1974

유산염배지(乳酸鹽培地)에서 배양(培養)한 Selenomonas ruminantium 균체(菌體)를 초음파처리후(超音波處理後) 추출(抽出)한 효소(酵素)와 cofactor로서 ATP, CoA, $Mg^{++}$, UTP를 사용(使用)하여 in vitro에서 당지질(糖脂質)을 생합성(生合成)하여 본 결과(結果)는 다음과 같다. 1. Selenomonas ruminantium의 효소(酵素)에 의(依)한 $^{14}C-glucosamine$과 tridecyl-Co A로 부터의 당지질(糖脂質)의 in vitro 생합성(生合成)은 ATP, $Mg^{++}$, UTP,에 의(依)하여 촉진(促進)되었다. 2. 효소추출액(酵素抽出液)을 원심분리(遠心分離)로 분획(分劃)하고 각분획(各分劃)의 glycolipid 합성능(合成能)을 본 결과(結果) 105,000의 supernatant에 6,000rpm supernatant의 약(約) 2배(倍)의 효소활성(酵素活性)을 나타내는 조효소(粗酵素)가 존재(存在)함을 알았다. 3. 효소반응(酵素反應)은 30분(分)일 때 대부분(大部分)이 진행(進行)되며 1시간(時間)에서 거의 완결(完結)됨을 알았다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권23호
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• pp.10469-10473
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• 2015

Background: Solanum nigrum L. has been used in traditional Chinese medicine because of its diuretic and antipyretic effects. The present research concerned effects of crude polysaccharides isolated from Solanum nigrum L. on erythrocyte membranes of tumor-bearing $S_{180}$ and $H_{22}$ in mice. Materials and Methods: Fluorescence-labeled red blood cell membranes were used with DPH fluorescence spectrophotometry to examine erythrocyte membrane fluidity, and colorimetry to determine degree of erythrocyte surface membrane blocking. Extent of reaction by tumor-bearing mice with the enzyme erythrocyte membrane bubble shadow detection of red cell membrane variation in the degree of closure before and after administration. Results: Solanum nigrum polysaccharide could significantly improve the $S_{180}$ and $H_{22}$ tumor-bearing mice erythrocyte membrane fluidity, compared with the control group, the difference was significant (p<0.01), SNL can significantly improve the red blood cell membrane and then $S_{180}$ tumor-bearing mice sealing ability, compared with the negative control group, the difference was significant(p<0.05, p<0.01). $H_{22}$ tumor-bearing mice can increase red cell membrane and then sealing ability, the difference was significant (p<0.05). Solanum nigrum polysaccharide degree of fluidity and blocking two transplanted tumors in mice restored the ability to raise the red cell membrane has a significant effect. Conclusions: Solanum nigrum L.-type mice transplanted tumor can affect the red blood cell membrane fluidity and re-closed, through the red cell membrane of red blood cells to enhance the immune function of the possibility of erythrocyte immunity against tumor formation garland provide experimental basis.

• Asian Pacific Journal of Cancer Prevention
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• 제15권1호
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• pp.475-481
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• 2014

Background: Pereskia sacharosa is a genus of cacti widely used in folk medicine for cancer-related treatment. Anti-proliferative effects have been studied in recent years against colon, breast, cervical and lung cancer cell lines, with promising results. We here extended study of anti-proliferative effects to a blood malignancy, leukemia. Materials and Methods: Two leukemic cell lines, MV4-11 (acute myeloid leukemia) and K562 (chronic myeloid leukemia), were studied. $IC_{50}$ concentrations were determined and apoptosis and cell cycle regulation were studied by flow cytometric analysis. The expression of apoptosis and cell-cycle related regulatory proteins was assessed by Western blotting. Results: P sacharosa inhibited growth of MV4-11 and K562 cells in a dose-dependent manner. The mode of cell death was via induction of intrinsic apoptotic pathways and cell cycle arrest. There was profound up-regulation of cytochrome c, caspases, p21 and p53 expression and repression of Akt and Bcl-2 expression in treated cells. Conclusions: These results suggest that P sacharosa induces leukemic cell death via apoptosis induction and changes in cell cycle checkpoint, thus deserves further study for anti-leukemic potential.

• 한국미생물·생명공학회지
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• 제11권1호
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• pp.23-32
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• 1983

본 연구에서는 Aspergillus niger KG79에서 두종류의 D-xylanase를 분리.정제하여 그 특성을 규명하였다. 이 두 종류의 D-xylanase의 물리화학적 및 동력학적 특성은 큰 차이가 없었다. 이들 Xylanase는 D-Xylan으로부터 Xylose, Xylobiose와 Xylotriose를 분해 생성하였다. 그러나 보리짚 Xylan을 기질로 사용했을 경우에는 Xylanase I 은 II보다 측쇄 arabinose를 상당히 빨리 분해하였다. 이들 효소에 의한 Xylan의 분해도는 기질의 종류에 따라 차이가 나서 보리짚 Xylan과 larchwood Xylan의 분해도는 각각 10%와 25%(환원당량) 정도로 나타났다. 순수정제된 Xylanase와 $\beta$-Xylosidase를 사용하여 재조합한 Xylanase계의 기질분해력을 비교한 결과 최적조건에서 보리짚과 larchwood Xylan은 각각 28%와 54%씩 분해 전환되었다. 이러한 결과는 Xylan의 효소분해도의 제한요소는 기질의 물리적 특성의 차이에 기인함을 추정할 수 있었다.

• 한국미생물·생명공학회지
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• 제11권1호
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• pp.47-52
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• 1983

Aspergillus sp.C-58 균주가 생산하는 extracellular inulase에 대하여 pH, charcoal처리 및 ammonium sulfate로 분별염석한 후 DEAE- cellulose를 이용한 column chromatography에 의하여 3개의 효소단백질(Peak I, II, III)로 분획되었으며 그 비율은 31. 1 : 1.7 : 1이였다. P- I, II의 I/S는 그 비율이 0.23 및 0.24로 거의 동일하였으나 P-III는 1.1로 P-I및 P-II와 상이하였다. Peak I 효소에 대하여 DEAE-Sephadex A-50을 이용한 ion exchange chromatography에 의하여 추출효소에 비교하여 약 408배 정제되었으며 다시 Sephadex G-75 및 Sephadex G-100에 2회 gel filtration하여 약 482배 정제되었다. 이상과 같이 정제한 Peak I의 효소액은 poly acrylamide를 이용한 disc gel electrophoresis 및 ultra centrifugation에 의하여 단일 단백질로 확인되었다.

• 한국수산과학회지
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• 제21권2호
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• pp.85-96
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• 1988

Purification and some properties of alkaline proteinases in the pyloric caeca of skipjack, Katsuwonus vagans, were investigated. Four alkaline proteinases, temporarily designated proteinases I, II, III and IV, were identified from the tissue extract of the pyloric caeca by ammonium sulfate fractionation, DEAE-Sephadex A-50 chromatography, and Sephadex G-100 and G-200 gel filtration. Result of disc-polyacrylamide gel electrophoretic analysis showed that the purified proteinases II and III were homogenous with the yields of $1.5\%\;and\;1.2\%$, and those specific activities were increased to 33 to 37 fold over that of the crude enzyme solution, respectively. Molecular weight of the proteinases II and III determined by sephadex G-100 gel filtration were 28,500 and 24,200, respectively. The optimum conditions for the caseinolytic activity of the two enzymes were pH 9.6 and $48^{\circ}C$. The reaction rates of the two alkaline proteinases were constant to the reaction time to 80 min in the reaction mixture of $3.4{\mu}g/ml$ of enzyme concentration and $2\%$ casein solution. The Km values against casein substrate determined by the method of Lineweaver-Burk were $0.56\%$ for proteinase II and $0.30\%$ for proteinase II. The proteinases II and III were inactivated under the presence of $Ag^+,\;Hg^{2+},\;Ni{2+},\;Fe^{2+},\;and\;Cu^{2+}$, and but activated by $Mn^{2+}\;and\;Ca^{2+}$ and markedly inhibited by the soybean trypsin inhibitor and N-p-toluenesulfonyl-L-lysine chloromethyl ketone. Therefore, the proteinases II and III were found to be a group of serine proteases and assured to be trypsin-like proteinases.

• 한국수산과학회지
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• 제53권3호
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• pp.308-315
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• 2020

To obtain a value-added product from the non-toxic brown-backed toadfish Lagocephalus gloveri (pufferfish), we developed a retort pouched pufferfish soup (RPS) and characterized its processing conditions and quality metrics. We found that the most appropriate manufacturing process for the RPS consisted of detoxifying and cold-water dipping the pufferfish flesh, blanching it, and adding it to the retort pouch along with other ingredients (hot-water extract of pufferfish head and carcass, radish, bean sprouts, and garlic), after which the pouch was sealed, sterilized (120℃, F₀ value 7.5-10 min.), cooled, and inspected. The moisture, crude protein, and total volatile basic nitrogen contents of the RPS were 97.2%, 1.3% and 7.7 mg/100 g, respectively. The total free amino acid content was 903.2 mg/100 g, and the main free amino acids were glutamic acid, taurine, lysine, glycine, threonine, alanine, arginine, proline, and hydroxyproline. Sterilizing the RPS for up to F₀ value 10 min. did not cause any major problems in terms of chemical or sensory qualities. This RPS has good storage stability and organoleptic qualities compared with similar commercial pufferfish soups and is suitable for commercialization as a value-added instant seafood soup.

• Korean Chemical Engineering Research
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• 제52권4호
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• pp.497-502
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• 2014

바이오매스 유래 타르 및 왁스 성분은 파클리탁셀의 분리정제에 많은 악영향을 미치기 때문에 반드시 제거되어야 한다. 흡착제 처리는 식물세포배양으로부터 유래된 타르 및 왁스 성분을 매우 간단하고 효과적으로 제거할 수 있는 방법이다. 본 연구에서는 전처리 단계에서 상용흡착제 실로퓨트를 이용한 흡착제 처리 공정에서 주요 공정 변수들(온도, 시간, 용매, 건조시료/실로퓨트 비율)을 최적화하였다. 최적의 흡착제 처리 온도, 시간, 용매, 건조시료/실로퓨트 비율은 각각 $30^{\circ}C$, 15분, 메틸렌 클로라이드, 1:1(w/w)이었다. 이러한 결과는 TGA와 HPLC 분석을 통한 유기물의 흡착 양상으로도 확인하였다. 흡착제 처리 단계에서 순도 증가는 미미하였으나 타르 및 왁스 성분의 제거로 인한 후속공정의 공정 가능성과 편리성에 상당히 영향을 미쳤다.

• 한국토양비료학회지
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• 제49권4호
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• pp.401-408
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• 2016

The present study described the isolation of 2-methylbutyric acid (2-MBA) produced from Bacillus pumilus L1, to subsequently investigate its nematicidal activity for the control of the root-knot nematode. The results showed that 2-MBA could be purified by chromatographic techniques and was identified using nuclear magnetic resonance and liquid chromatography-mass spectrometry. Crude extract and partially purified compounds had a significant effect on the inhibition of egg hatchability and second-stage juvenile (J2) mortality. A dose-dependent effect of 2-MBA was observed for J2 mortality and egg hatchability. Egg hatchability was 69.2%, 59.9%, 32.7%, and 0.0% at 125, 250, 500, and $1000{\mu}g\;mL^{-1}$ of 2-MBA after 4 d of incubation, respectively. Meanwhile, J2 mortality was in the range of 24.4%-100.0% after 2 d of incubation, depending on the concentrations of 2-MBA used. A pot experiment also demonstrated that treatment of B. pumilus L1 culture caused a significant reduction in the number of galls, egg masses, and J2 population than that of the tap water (TW) control. However, as the B. pumilus L1 culture concentration was decreased, the efficacy of nematode control by treatment of B. pumilus L1 culture was reduced compared to that of TW. B. pumilus L1 inoculation at different concentrations also promoted cucumber plant growth. Therefore, our study demonstrated the potential of 2-MBA from B. pumilus L1 as a biocontrol agent against the root-knot nematode and a plant growth promoter for cucumber plants.

• 미생물학회지
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• 제32권1호
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• pp.78-83
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• 1994

초산 생성 균주인 Acetobacter sp. HA로부터 membrane-bound alcohol dehydrogenase(ADH)를 분리 정제하였다. 세포막 분획과 세포막에서 효소의 용출 그리고 크로마토그라피 방법 등을 적용하여 수율 35%, 153배 정제된 효소를 획득하였다. 정제된 효소의 분자량은 330,000 dalton이었으며, 각각 분자량 79,000과 49,000 그리고 45,000 dalton을 가진 세 종류의 subunit로 이루어져 있었다. 그리고 흡수 스펙트럼의 분석 결과 본 효소에는 cytochrome c가 존재함을 확인할 수 있었다. 본 효소는 메탄올을 제외한 1차 지방족 알코올을 기질로 이용할 수 있었으며, formaldehyde, acetaldehyde 그리고 glutaraldehyde의 경우에도 다소간 기질로 이용될 수 있었다. 본 효소의 에탄올에 대한 $K_m$값은 1.38mM이었으며, 최적 pH와 온도는 각각 5.0~6.0과 32${\circ}C$이었다. 본 효소의 활성은 $V_2O_5$와 $ZnCl_2,\; NiCl_2$같은 금속 이온에 의하여 저해되었다.