• 한국미생물·생명공학회지
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• 제19권3호
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• pp.235-241
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• 1991

Streptomyces melanosporofaciens로 동정된 strain 88-GT-161 균주는, 그람양성세균 및 식물병원성 곰팡이에 각각 항균활성을 나타내는 phthalate 유도체 및 염기성 마크로라이드의 새로운 항생물질을 생산하는 것으로 밝혀졌다. 이들 활성물질들의 분리. 정제과정에서 in vitro 및 in vivo(포트 시험) 항균활성을 조사하였으며, phtahalate 유도체 항생물질은 IR, NMR, 질량분석 스펙트럼 조사를 통하여 bis(2-ethylexyl) phthalate(dioctyl phthalate)로 동정하였다. Dioctyl phthalate가 Streptomyces sp.에 의하여 생산되며 생합성된 이 화합물이 항균활성을 가진다는 사실이 보고된 것은 이것이 처음이다.

• 생약학회지
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• 제53권3호
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• pp.125-132
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• 2022

In this study, we investigated anti-oxidative and anti-inflammatory efficacy, and identified their constituents from Brassica napus L. (Korean name: Yuchae) whole plant. Upon the anti-oxidative activities screening, the ethanol extract exhibited potent DPPH and ABTS⁺ radical scavenging activities. On the anti-inflammation studies using LPS-induced RAW264.7 cells, the extract inhibited the production of NO and pro-inflammatory cytokines (TNF-α, IL-1β, IL-6) effectively. To identify major constituents of B. napus extract, further purification was performed and led to isolation of two compounds; isorhamnetin 3,7-O-diglucoside(1) and isorhamnetin 3-O-glucoside(2). Quantitative analysis by high pressure liquid chromatography (HPLC) determined the flavonoid 1 as the major constituent. Isolated compounds showed DPPH radical scavenging effects and decreased NO levels without causing cell toxicities. These results indicate that the extract of Yuchae, a rich plant resource in Jeju Island, could be potentially applicable as an anti-oxidative and/or anti-inflammatory ingredients.

• 약학회지
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• 제50권1호
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• pp.21-25
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• 2006

Reiterated normal-phase column chromatography lead to the isolation and purification of six known compounds but for the first time from the whole plant of Sasa borealis (Hack.) Makino (Gramineae): tricin 4'-O-(erythro-${\beta}$-guaia-cylglyceryl) ether (1), tricin 4'-O-(threo-${\beta}$-guaiacylglyceryl) ether (2), tricin 4'-O-[erythro-${\beta}$-guaiacyl-(9'-O-acetyl)-glyceryl] ether (3), tricin 4'-O-[threo-${\beta}$-guaiacyl-(9'-O-acetyl)-glyceryl] ether (4), (-)-pinoresinol (5), and vanillin (6). The structures of the compounds (1-6) were established based on interpretation of high resolution NMR (COSY, HSQC, HMBC, and NOESY) spectral data. In particular, compounds 1 and 3 were diastereomers of compounds 2 and 4, respectively. These two sets of diastereomers were able to be simultaneously identified and quantified by a gradient reversed-phase HPLC method with UV photodiode array, This sensitive HPLC method is noteworthy as a simultaneous separation and identification method to test the extract of the family Gramineae which contains these compounds.

• Fisheries and Aquatic Sciences
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• 제24권4호
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• pp.163-170
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• 2021

Peptides are naturally occurring biological molecules that are found in all living organisms. These biologically active peptides play a key role in various biological processes. The aim of this study is the extraction and the purification of bioactive materials that induce relaxation of an apical muscle from the pyloric caeca of Patiria pectinifera. The acidified pyloric caeca extract was partially separated by the solid phase extraction using a stepwise gradient on Sep-Pak C18 cartridge. Among the fractions, materials eluted with 60% methanol/0.1% trifluoroacetic acid was put a thorough of a series of high performance liquid chromatography (HPLC) steps to isolate a neuropeptide with relaxation activity. The purified compound was eluted at 28% acetonitrile in 0.1% trifluoroacetic acid with retention time of 25.8 min on the CAPCELL-PAK C18 reversed-phase column. To determine the molecular weight and the amino acid sequence of the purified peptide, LC-MS and Edman degradation method were used, respectively. The primary structure of the peptide was determined to be FGMGGAYDPLSAGFTD which corresponded to the amino acid sequence of a starfish myorelaxant peptide (SMP) isotype (SMPb) found in the cDNA sequence encoding SMPa and its isotypes. In this study, a muscle relaxant neuropeptide (SMPb) has been isolated from pyloric caeca of starfish P. pectinifera. This is the first report of SMPb isolation on the protein level from P. pectinifera.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2011년도 임시총회 및 추계학술발표회
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• pp.20-20
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• 2011

In this study, we analyzed 80%MeOH extract of fruits of sorbaria sorbifolia var. stellipila MAX. to measure the total antioxidant capacity by oxygen radical absorbance capacity (ORAC) assay, individual flavonoid content by high-performance liquid chromatography (HPLC). n-Hexane ($1.02{\pm}0.036$), $CH_2Cl_2$ ($0.95{\pm}0.025$), EtOAc ($1.94{\pm}0.065$), n-BuOH ($1.98{\pm}0.054$), D.W. ($1.2{\pm}0.032$) fractions were examined antioxidative activity by ORAC assay. It was revealed that EtOAc($1.94{\pm}0.065$), n-BuOH($1.98{\pm}0.054$) fractions had significant antioxidative activity. The isolation and separation were facilitated using open column chromatography, while separation, purification and identification were accomplished by using high-performance liquid chromatography (HPLC) and nuclear magnetic resonance (NMR).

• Parasites, Hosts and Diseases
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• 제56권1호
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• pp.25-32
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• 2018

Molecular techniques have been introduced for malaria diagnosis because they offer greater sensitivity and specificity than microscopic examinations. Therefore, DNA isolation methods have been developed for easy preparation and cost effectiveness. The present study described a simple protocol for Plasmodium DNA isolation from EDTA-whole blood. This study demonstrated that after heating infected blood samples with Tris-EDTA buffer and proteinase K solution, without isolation and purification steps, the supernatant can be used as a DNA template for amplification by PCR. The sensitivity of the extracted DNA of Plasmodium falciparum and Plasmodium vivax was separately analyzed by both PCR and semi-nested PCR (Sn-PCR). The results revealed that for PCR the limit of detection was $40parasites/{\mu}l$ for P. falciparum and $35.2parasites/{\mu}l$ for P. vivax, whereas for Sn-PCR the limit of detection was $1.6parasites/{\mu}l$ for P. falciparum and $1.4parasites/{\mu}l$ for P. vivax. This new method was then verified by DNA extraction of whole blood from 11 asymptomatic Myanmar migrant workers and analyzed by Sn-PCR. The results revealed that DNA can be extracted from all samples, and there were 2 positive samples for Plasmodium (P. falciparum and P. vivax). Therefore, the protocol can be an alternative method for DNA extraction in laboratories with limited resources and a lack of trained technicians for malaria diagnosis. In addition, this protocol can be applied for subclinical cases, and this will be helpful for epidemiology and control.

• 한국산림과학회지
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• 제74권1호
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• pp.29-36
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• 1986

이태리포푸라 I-214 (Populus euramericana cv. I-214)의 기내배양(器內培養)한 엽육조직(葉肉組織)에서 원형질체(原形質體) 분리(分離)에 미치는 몇가지 요인(要因)에 대(對)해 조사(調査), 검토(檢討)하였다. 기내(器內)에서 배양(培養)된 아(芽)를 다량(多量)으로 증식(增植)하기 위한 배지(培地)는 MS 기본배지(基本培地)에 $0.1mg/{\ell}$의 BAP를 첨가(添加)한 것이 가상 좋은 성적을 나타냈다. 엽(葉) 1g 당(當) $2.4{\times}10^6$개의 가장 높은 원형질체(原形質體) 분리(分離) 빈도를 나타낸 것은 Cellulase R-10 2 %, Macerozyme R-10 0.8 %, Hemicellulase 1.2 %, Driselase 2.0 %, Pectolyase Y-23 0.05 %에 DTT와 MES 완충액을 첨가(添加)한 후 삼투압 안정제로 0.6 M의 Mannitol을 넣고 pH를 5.6으로 조정한 효소용액(酵素溶液)이었다. CPW 용액(溶液)으로 세정(洗淨)한 후 0.6 M의 Sucrose 용액(溶液)에 처리(處理)한 것이 회수율(回收率) 51.8 %로 가장 높게 나타났다.

• BMB Reports
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• 제37권1호
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• pp.59-74
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• 2004

The study of whole patterns of changes in protein expression and their modifications, or proteomics, presents both technological advances as well as formidable challenges to biological researchers. Nutrition research and the food sciences in general will be strongly influenced by the new knowledge generated by the proteomics approach. This review examines the different aspects of proteomics technologies, while emphasizing the value of consideration of "traditional" aspects of protein separation. These include the choice of the cell, the subcellular fraction, and the isolation and purification of the relevant protein fraction (if known) by protein chromatographic procedures. Qualitative and quantitative analyses of proteins and their peptides formed by proteolytic hydrolysis have been substantially enhanced by the development of mass spectrometry technologies in combination with nanoscale fluidics analysis. These are described, as are the pros and cons of each method in current use.

• 미생물학회지
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• 제19권1호
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• pp.31-37
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• 1981

Avicelase, CMCase and salicinase of A.phoenicis K.U. 175 were purified from wheat bran culture by salting out with ammonium sulfate, dialysis and successive column chromatography Sephadex G-100. Optimum pH and temperature of avicelase were pH 3.8-4.8, $35-55^{\circ}C$ and that of CMCase, salicinase were pH4.5-5.5, $45-60^{\circ}C$ and pH 4.5-6.0, $45-60^{\circ}C$ respectively. These enzymes were relatively thermostable, alkali unstable and inhibited by $Ca^{++},\;Mn^{++},\;Cu^{++},\;and\;Hg^{++}$. Km values of avicelase, CMCase and salicinase were calculated to be $1.5{\times}10^{-4}M,\;5.5{\times}10^{-4}M\;and\;2.75{\times}10^{-5}M$ and Vmax values $1.66{\times}10^{-4}mM/min,\;3.33{\times}10^{-4}mM/min\;and\;1.14{\times}10^{-4}mM/min$, respectively.

• Journal of Microbiology and Biotechnology
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• 제10권4호
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• pp.539-543
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• 2000

A new bacteriocin, named lacticin 10790, was purified from Lactococcus lactis subsp. cremoris KFCC 10790 by sequential adsorption, immobilized metal-affinity, cation-exchange, and $C_{18}$ reverse-phase chromatographies. The molecular mass of the bacteriocin was estimated to be between 3,000 and 3,500 Da. Lacticin 10790 showed a broad antimicrobial spectrum against many gram-positive bacteria. The bacteriocin was stable to heat and in the pH range between 2 and 6. Lacticin 10790 was destroyed by digestion with proteases and exhibited a bactericidal mode of action. An amino acid composition analysis of purified lacticin 10790 revealed a high concentration of hydrophobic amino acids. The N terminus of the bacteriocin was found to be blocked, upon analysis by Edman degradation. The results suggest that lacticin 10790 is a class I bacteriocin.