• Applied Biological Chemistry
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• 제22권1호
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• pp.10-17
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• 1979

인삼엽(人蔘葉)을 ether, benzene-butanol 혼합용매등으로 색소류(色素類)기타 지용성물질(脂溶性物質)등을 제거하고, methanol과 n-butanol로 연속추출하여 n-butanol soluble saponin을 얻고 계속해서 liquid chromotography 를 행하여 분획별(身劃別)로 인삼엽(人蔘葉) saponin을 분리 정제하였고, 그 화학적(化學的)및 분자구조학적(分子構造學的) 특성(特性)을 인삼근(人蔘根) saponin과 비교 검토한 결과는 다음과 같다. 1. 인삼근(人蔘根)과 엽(葉) saponin의 2차원(차원) TLC pattern은 다소(多少) 차이(差異)차가 있으나, 다수(多數)의 spot들이 일치(一致)하였다. 인삼엽(人蔘葉)에는 ginsenoside Rd가 존재하지 않고, ginsenoside Rc는 엽(葉)에, ginsenoside $Re_1$ 및 $Rg_1$는 근부(根部)에 더 많이 분포(分布)되어 있었다. 2. 인삼엽(人蔘葉)에는 인삼근(人蔘根)에 비하여, panaxadiol 계(系) saponin이 panaxatriol 계(系) saponin보다 더높은 함량을 보였다. 3. n-butanol soluble saponin으로 liquid chromatography를 행하여 인삼근(人蔘根)에서는 Fr. 1.과 Fr. 2.를 얻었고, 인삼엽(人襲葉)에서는 Fr. 1, Fr. 2. 및 Fr. 3.의 분획(分劃)을 얻을 수 있었다. 인삼근(人蔘根)의 Fr. 1 및 엽(葉)의 Fr. 1.과 Fr. 2.등 3매의 분획(分劃)은 saponin test반응(反應)에 양성(陽性)을 보이고, 그 IR spectra는 동일(同一)한 것으로 나타났다.

• 생명과학회지
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• 제15권1호
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• pp.94-99
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• 2005

충청남도 목천과 충청북도 오창 근교의 토양으로부터 망간을 산화하는 64 집락을 분리하고 이 중 망간 산화능이 가장 우수한 한 균주를 최종 선별하여 생리, 생화학적 특성을 조사하고, 16S rRNA 염기 서열분석 등을 통하여 동정한 결과 최종 선별된 균주는 Aeromonas sp. MN44로 확인되었다. 최종 선별된 Aeromenas sp. MN44는 lactose를 제외한 여러 당들은 이용하지 못하였으며, 중금속내성은 lithium과 manganese에 대해서는 mg/ml 단위의 높은 농도까지 중금속 내성을 가지고 있었지만 cadmium에는 전혀 내성을 나타내지 않았다. 또 kanamycin, chloramphenicol, ampicillin, tetracycline, spectinomycin등 조사한 모든 항생제에 대해 전혀 내성을 갖지 않았다. Aeromonas sp. MN44가 생성하는 망간산화물질의 최적 pH는 pH 7.4로 확인되었으며, 이 균이 생성하는 망간 산화 factor는 proteinase K와 가열처리에 의해 저해되는 단백질이고, ammonium sulfate 침전과 ion exchange chromatography 그리고 gel filtration의 단계를 통해 부분 정제한 망간 산화 factor의 분자량은 약 113 kDa로 확인되었다.

• Bulletin of the Korean Chemical Society
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• 제32권7호
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• pp.2418-2422
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• 2011

The enantiomeric separation of lactic acid for its absolute configuration has become important task for understanding its biological origin and metabolic process involved in the formation of lactic acid. It involves the conversion of enantiomers as diastereomeric O-pentafluoropropionylated (S)-(+)-3-methyl-2-butyl ester and the direct separation by gas chromatography-mass spectrometry on a achiral capillary column. The (R)- and (S)-lactic acids were completely separated with a high resolution of 1.9. The newly developed method showed good linearity (r ${\geq}$ 0.999), precision (% relative standard deviation = 3.4-6.2), and accuracy (% relative error = -7.7-1.4) with the detection limit of 0.011 ${\mu}g/mL$. When the method was applied to determine the absolute configuration of lactic acid in Lactobacillus delbrueckii subsp. lactis 304 (LAB 304), the composition (%) of (R)-lactic acid in the cell pellet and in the culture medium were $89.0{\pm}0.1$ and $78.2{\pm}0.4$, respectively. Thus, it was verified that the present method is useful for the identification and composition test of lactic enantiomers in microorganisms.

• 약학회지
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• 제41권5호
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• pp.615-621
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• 1997

Dehydropeptidase-I (DHP-I) was solubilized from porcine kidney by treatment with n-butanol and partially purified 19.25 fold by $(NH_4)_2SO_4$ precipitation, DEAE-Sepharose CL-6B ion exchange chromatography and Sephacryl S-300 HR chromatography with an overall yield of 19.16. DHP-I showed its optimal activity at pH 7.5 and 25$^{\circ}C$. Its activity was stable under neutral and alkaline conditions, but was disappeared under acidic condition. And DHP-I was heat-labile and its activity remained at 45$^{\circ}C$ for 3hrs. The enzyme was not inhibited by dicationic ions, while its activity was increased by $Co^{2+}$(1mM) and $Zn^{2+}$ (0.1mM). The enzyme was inhibited by EDTA and N-ethylmaleimide. The relative molecular mass of DHP-I was estimated to be approximately 100kDa by gel filtration chromatography. The $K_m$ value of DHP-I for glycyldehydrophenylalanine (GDHP) was 1.98mM. DWP20418 [(1R, 5S, 6S)-6-[1-(R)-Hydroxyethyl]-1-methyl-2-[(2S, 4S)-2-(piperazinylcarbonyl)-1-(R)-hydroxyethyl)pyrrolidine-4-thio]carbapen-2-em-3-carboxylic acid], compared with meropenem (MEPM), was rather easily hydrolized by DHP-I, while it was four times more resistant than imipenem (IPM) to DHP-I.

• Bulletin of the Korean Chemical Society
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• 제31권12호
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• pp.3567-3572
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• 2010

Black pine barks from the southern region of Korea were extracted using pressurized hot water and the water soluble extracts were then separated in a stepwise fashion using a variety of solvents, column chromatography (CC), thin layer chromatography (TLC), and high pressure liquid chromatography (HPLC). The antioxidant activities of each fraction and the active compounds were determined based on the radical scavenging activities of 2,2-diphenyl-1-picrylhydrazyl (DPPH), reductive potential of ferric ion, and total phenol contents. A DPPH test showed that the half maximal effective concentration ($EC_{50}$ value : $6.59{\pm}0.31\;{\mu}g/mL$) of the ethyl acetate fraction (ca. 0.67%) was almost the same as that of the control compounds and inversely proportional to the value of the total phenol contents. The cell viability of the water extracts was confirmed by methyl thiazol-2-yl-2,5-diphenyl tetrazolium bromide (MTT) with enzyme linked immune sorbent assay (ELISA). Catechin, epicatechin, quercetin and ferulic acid were isolated from the ethyl acetate fraction as active compounds and identified by nuclear magnetic resonance. The antioxidant activity as value of DPPH of each of the separated compounds was lower than the ethyl acetate fraction, and ferulic acid was the lowest among these compounds.

• Applied Biological Chemistry
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• 제41권7호
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• pp.522-526
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• 1998

콩 유용성분 탐색의 일환으로 그리고 향후 콩 ferritin 항체 및 유전자 확보를 목표로 발아된 콩으로부터 ferritin을 분리 정제하여 그 몇가지 특성을 조사하였다. 72시간 발아된 콩으로부터 ammonium sulfate 침전(0.55 saturation), DEAE-cellulose, Sephacryl S-300, Bio-Scale Q2 column chromatographies를 통하여 ferritin을 분리하였다. 정제된 콩 ferritin은 SDS-PAGE 분석에서 21 kDa의 크기를 나타냈으며, Sephacry S-300을 통한 겔거르기 chromatography와 non-denaturing 폴리아크릴아마이드 전기영동 분석에서 $510{\sim}560\;kDa$의 크기로 측정 되었다. 또한, immunodiffusion test에서 anti-soybean ferritin antiserum과 상호 반응하였다. 원자흡광광도계와 표준 철 용액을 이용한 정제된 콩 ferritin 중의 철 함유량은 833 mol Fe/mol protein 이었으며, 이는 호박씨로부터 분리한 ferritin보다 31배 더 많은 양의 철 함유량 이었다. 정제된 콩 ferritin중의 철은 horse spleen ferritin 중의 철과 유사하게 iron staining 되었다.

• 한국자원식물학회지
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• 제32권4호
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• pp.264-269
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• 2019

쉽싸리(L. lucidus)가 가지는 효능의 하나로 알려진 항염증효과의 활성물질을 파악하기 위하여 본 연구를 수행하였다. 항염효과는 LPS로 활성화한 macrophage 264.7이 생산하는 NO의 감소효과를 측정함으로써 평가하였다. 쉽싸리 추출물에서 얻은 비극성 분획물인 $CHCl_3$ 분획물은 농도의존적으로 현저히 NO 생산을 감소시켰다. 이에 비해 극성 분획물인 BuOH 분획물은 그 효과가 약하였다. Silica gel column chromatography에 의해 이 $CHCl_3$ 분획물로부터 주요 화합물인 ursolic acid를 분리하고 분광학적 방법으로 동정할 수 있었다. 효과가 약하였던 BuOH 분획물로부터 diaion HP-20 column chromatography와 sephadex LH-20 column chromatography로 이 분획의 주요 화합물인 rosmarinic acid를 분리하고 역시 분광학적 방법으로 동정하였다. Ursolic acid는 농도의존적으로 NO 생산을 억제하였으나 rosmarinic acid는 그 효과가 상대적으로 약하였다. 이러한 사실로부터 쉽싸리의 항염효과는 주로 ursolic acid의 존재 때문임을 알 수 있었다.

• FOCUS: LIFE SCIENCE
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• 제1호
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• pp.1.1-1.4
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• 2024

This article provides comprehensive guidance on the maintenance, cleaning, regeneration, and storage of silica-based HPLC (High-Performance Liquid Chromatography) columns. The general considerations emphasize the importance of using in-line filters and guard cartridges to protect columns from blockage and irreversible sample adsorption. While these measures help, contamination by strongly adsorbed sample components can still occur over time, leading to an increase in back pressure, loss of efficiency, and other issues. To maximize column lifetime, especially with UHPLC (Ultra-High Performance Liquid Chromatography) columns, it is advisable to use ultra-pure solvents, freshly prepared aqueous mobile phases, and to filter all samples, standards, and mobile phases. Additionally, an in-line filter system and sample clean-up on dirty samples are recommended. However, in cases of irreversible compound adsorption or column voiding, regeneration may not be possible. The document also provides specific recommendations for column cleaning procedures, including the flushing procedures for various types of columns such as reversed phase, unbonded silica, bonded normal phase, anion exchange, cation exchange, and size exclusion columns for proteins. The flushing procedures involve using specific solvents in a series to clean and regenerate the columns. It is emphasized that the flow rate during flushing should not exceed the specified limit for the particular column, and the last solvent used should be compatible with the mobile phase. Furthermore, the article outlines the storage conditions for silica based HPLC columns, highlighting the impact of storage conditions on the column's lifetime. It is recommended to flush all buffers, salts, and ion-pairing reagents from the column before storage. The storage solvent should ideally match the one used in the initial column test chromatogram provided by the manufacturer, and column end plugs should be fitted to prevent solvent evaporation and drying out of the packing bed.

• 한국미생물·생명공학회지
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• 제25권1호
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• pp.62-67
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• 1997

A potential biocontrol bacterium, YB-70 was isolated from a rhizosphere in suppressive soil and identified as a strain of Bacillus subtilis. In several biochemical and in vitro antibiosis tests on Fusarium solani with the culture filterates from B. subtilis YB-70, we found that antifungal mechanism of B. subtilis YB-70 was mediated by antibiotic substances produced from the bacterium. These antifungal substances were appeared to be hear-resistant, micromolecular, and ethy alcohol soluble. Antifungal agents produced by B. subtilis YB-70 showed strong inhibified against root-rotting fungi F. solani in in vivo pot test. An antifungal substance. YBS-1s, was purified from the culture broth of B. subtilis YB-70 by isoelectronic precipitation, silica gel column chromatography and Sephadex LH-20 column chromatography analysis by Fab-MASS, $^{1}$H-NMR, $^{13}$C-NMR, DEPT, and amino acid analyzer revealed that the YBS-1A was a peptide antibiotics of iturin class containing seven amino acids from five different groups, and the other(YBS-1B) was an analogue of iturin group composed of 11 amino acids with larher molecular weight of about 1, 500 dalton, which was lager than that of iturin A.

• 생명과학회지
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• 제12권4호
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• pp.483-489
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• 2002

S. cerevisiae에서 glucoamylase 유전자의 promoter와 signal sequence 그리고 MF$\alpha$1의 prosequence를 이용하여 합성 곤충 defensin를 발현하고 항균활성을 보유한 형태로 분비하는데 성공하였다. Defensin의 여러 생화학적인 특성을 조사한 결과 열 안정성이 높아 10$0^{\circ}C$에서 30분간 가열하여도 항균활성을 온전히 유지하였으며 조사한 pH 영역, 2.0-12.0에서 항균활성의 변화가 없었다. 또한 여러 단백질 분해효소를 처리하면 항균활성이 완전히 사라졌으나 전분질 분해효소, 섬유소분해효소 및 지질분해효소의 처리는 항균 활성에 전혀 영향이 없었다. 황산암모늄침전, SP-Sepharose column cormatography, RP-HPLC 등의 조작을 통해 defensin을 순수한 형태로 정제하였으며 Tricine-SDS-PAGE를 통해 분자량이 약 4.0 kDa임을 확인하였고 정제한 defensin은 항균활성을 보유하였다.