• Plant Resources
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• 제8권2호
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• pp.145-154
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• 2005

The antimicrobial effect of each fraction after fractionation of an ethanol extract of leaf mustard was examined in terms of nucleic acid, chloroform, ethylacetate, and butanol. The ethylacetate fraction, which showed the strongest level of antimicrobial effect among the different ethanol extract fractions of leaf mustard, was isolated and purified using silica gel column chromatography and HPLC, respectively, to obtain a single antimicrobial substance called KLM-1. The antimicrobial effect of this substance was 10 times higher than that of the ethylacetate fraction. A further study is on the way to confirm the structure of the antimicrobial substance KLM-1 through LC/Mass and NMR.

• Food Science and Biotechnology
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• 제17권3호
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• pp.679-682
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• 2008

A lactic acid bacterium producing an antimicrobial substance against Escherichia coli O157:H7 was isolated from raw milk and identified as Lactobacillus amylovorus ME-1. In addition to E. coli O157 :H7, the antimicrobial substance also inhibited the growth of Bacillus cereus, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella typhimurium, Staphylococcus aureus, Streptococcus agalactiae, Streptococcus pyrogenes, and Yersinia enterocolitica. The antimicrobial substance was stable at pH 2-12 and $121^{\circ}C$ for 15 min and insensitive to proteinase K, protease, amylase, and catalase. Purification of the antimicrobial substance was conducted through methanol and acetonitrile/ethylacetate extraction, ultrafiltration with a 500 Da cutoff, thin layer chromatography (TLC) with silicagel 60, and high performance liquid chromatography (HPLC) with a $C_{18}$ reverse phase column. The ${\lambda}_{max}$ of the purified antimicrobial substance was determined as 192 nm by ultra violet (UV) scanning, while the molecular weight was estimated as 453 Da based on the mass spectrum. Accordingly, the current results suggest that the antimicrobial substance from the L. amylovorus ME-1 was not a bacteriocin, but rather a new non-proteinaceous substance distinct from acidophilin, acidolin, diacetyl, and reuterin.

• 한국식품위생안전성학회지
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• 제10권4호
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• pp.225.1-230
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• 1995

The ethanol extract from the root bark of Morus alba showed the strongest antimicrobial activity on the growth of almost all the tested microorganisms which were food-borne pathogens and food-related microorganisms. 1) In order to isolate and purify of antimicrobial substance extracted from the root bark of Morus alba, the antimicrobial substance from the ethanol extract which exhibited a strong antimicrobial activity was purified by solvent fractionation, silica gel column chromatography, TLC and HPLC. Among the fractions fractionated by 4 kind of solvents from the ethanol extract, the antimicrobial activity of ethyl acetate fraction had the strongest antimicrobial activity against B. subtilis. Unknown compound were isolated from the ethyl acetate fraction by silica gel column chromatography, TLC and HPLC and the compounds showed strong absorbance at 207, 217 and 285 nm, therefore, it was supposed to be a kinds of aromatic compound.

• 한국환경과학회지
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• 제11권1호
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• pp.33-40
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• 2002

An antimicrobial substance-producing microorganism was isolated from soil samples. Based of the taxonomic characteristics of its morphological, cultural, physiological properties and 16s rRNA sequence alignment, this microorganism was identified as Pseudomonas aeruginosa, and we named Pseudomonas aeruginosa EL-KM. The optimal culture condition for production of antimicrobial substance was 1% mannitol, 0.4% yeast extract, 0.5% Nacl, 0.2% $K_2SO_4$, 100$\mu$M $MgSO_4$.$7H_2O$, 10$\mu$M $CaCl_2$.$2H_2O$, 1$\mu$M $FeSO_4$.$7H_2O$, 1$\mu$M $MnSO_4$.$4-5H_2O$, initial pH 7 and 200 rpm at 3$0^{\circ}C$. The purification of the antimicrbial substance was performed by silica gel column chromatographys, and fraction with TLC $R_f$ 0.77 value represented good antimicrobial activity. The crude antimicrbial substance was stable within a pH range of 3-10 and temperature range of 4$^{\circ}C$-121$^{\circ}C$ autoclaving. This crude antibacterial substance acted as bacteriolytic agent against Vibrio cholerae non-Ol ATCC 25872, and also exhibited excellent properties, when the substance was demonstrated against many other gram-positive, gram-negative bacteria, yeast and fungi.

• 한국미생물·생명공학회지
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• 제22권6호
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• pp.700-706
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• 1994

Lactic acid bacteria in Kimchi fermentation were tested for inhibitory activity against Gram positive bacteria and Gram negative bacteria. The Lactobacillus brevis (KCCM 35464) was found to produce a antimicrobial substance. It showed relatively wide range of inhibition spectrum against gram positive and gram negative bacteria and maintained the inhibitory activity between pH 4.0 and pH 9.0. The antimicrobial substance was obtained in the stationary growth phase and was purified by gel chromatography. The inhibitory effect of the antimicrobial substance on sensitive bacterial strains was determined by filter paper test. The activity of antimicrobial substa- nce was stable at 75$\circ$C. On the basis of its electrophoretic pattern is SDS-PAGE, antimicrobial substance appeared as a single band of 59 KDalton.

• 한국식품위생안전성학회지
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• 제10권3호
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• pp.139-145
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• 1995

In order to develop a natural food preservative, the root bark of Morus alba was extracted with several solvents, and then antimicrobial activity was investigated. The optimum extracting condition for the antimicrobial substance from the sample, minimum inhibitory concentration (MIC) of the extracted substance against microorganisms were also examined. The antimicrobial activity of the ethanol extract form the sample was stronger than those of the extracts by the other solvents such as water, methanol. ethyl acetate and acetone. The optimum extracting condition for antimicrobial substance from the sample was shaking extraction twice for 5 hours at room temperature in case of 7 times of absolute ethanol added to the crushed root bark of Morus alba. The ethanol extract from the root bark of Morus alba had strong B. cereus, L. monocytogenes and S. aureus. Especially, Bacillus species was the most susceptible to the extracted substance. The ethanol extract showed antimicrobial activity against Gram negative bacteria(MIC, 160~1600 ug/ml) and yeasts(MIC, 1600 ug/ml) such as C. albicans and S. acidifaeciens. The extract also showed growth inhibition against molds such as A. niger, A. parasiticus, A versicolar and T. viride.

• 한국식품저장유통학회지
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• 제1권1호
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• pp.1-7
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• 1994

To investigate the effect of several bactericides and fungicides against fruit & vegetable decay grapefruit seed extract(GFSE) was tested in a dilution solution. GFSE was shown to be effective against decay and rind breakdown and to extend the shelf-life of fruits and vegetables during storage and tiansport. Antimicrobial efficiency of GFSE on a wide spectra of gram + & - bacteria, moulds and yeasts was demonstrated by the measurement of minimal inhibitory concentrations performed (Bacteria 10-500ppm, Fungi:250-1,000ppm, Yeasts:100-250ppm). GFSE was separated and extracted into water-soluble fraction, water-insoluble and non-dialyzed fraction to isolate the antimicrobial substances. The water-soluble fraction showed the most active antimicrobial effect. The antimicrobial substances were isolated by gas chromatography. As the result of the isolation using GC, Peak-D was found to be the antimicrobial compound in GFSE. The identification of the most antimicrobial substance was carried out by using GC-MS.

• 한국미생물·생명공학회지
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• 제42권4호
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• pp.393-401
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• 2014

청국장으로부터 Bacillus cereus에 대한 항균활성이 가장 높은 균주를 분리하여 형태학적, 생화학적 특성과 16S rDNA 염기서열 결정을 통해 Bacillus subtilis HH28으로 동정 및 명명하였다. B. subilis HH28의 생육시기에 따른 항균활성을 측정해 본 결과, 생육이 대수증식기인 9시간부터 생성되어 사멸기인 60시간에 가장 높은 활성(80 AU/ml)을 나타내었고 144시간(6일)까지 항균활성을 유지하였다. 항균물질의 정제는 황산암모늄 침전과 DEAE-sepharose fast flow, sephacryl S-200HR를 이용하였고, 19.7배의 정제도와 38.4%의 수율로 정제되었다. Tricine SDS-PAGE와 directed detection을 통해 항균물질의 분자량이 약 3,500 Da임을 알 수 있었다. 또한 본 연구의 항균물질을 B. cereus 뿐만 아니라 Listeria monocytogenes, Vibrio parahaemolyticus의 식중독 균에서도 우수한 항균활성을 나타내었고, 항균작용의 기작은 미생물을 사멸시키는 살균작용이였다. 또한 온도 안정성 실험에서 $40-80^{\circ}C$까지 안정했고, pH 안정성 실험에서는 pH 2-9까지 안정하여 비교적 온도와 pH에 안정하였다. 효소에 대한 영향 실험에서는 단백질 분해효소에 의해 항균활성이 실활되어 본 연구의 항균물질은 단백질성임을 알 수 있었다. 위와 같은 특성으로 보아 B. subtilis HH28이 생산하는 항균물질은 천연 식품 보존제 및 사료 보존제, 항생제 대체 의약품으로 사용할 수 있는 잠재력을 가지고 있으며, 향후 이 항균물질의 정확한 구조 및 특성 규명 등의 연구가 필요하다.

• 한국식품영양과학회지
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• 제24권5호
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• pp.695-701
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• 1995

The ethanol extract of leaf mustard(Brassica juncea) exhibiting high antimicrobial activities was fractionated in the order of hexane, chloroform, ethylacetate and butanol fractions to test antimicrobial activity. The highest antimicrobial activity for the bacteria tested was found in the ethylacetate fraction, but a lesser extent in the butanol fraction. In contrast to antimicrobial activity for the bacteria, both ethylacetate and butanol fractions showed weak antimicrobial activity for yeasts. Unknown compound A in the ethylacetate fraction which exhibited a strong antimicrobial activity was isolated by silica gel column chromatography and HPLC, and exhibited 9 times more antimicrobial activity than the ethylacetate fraction.

• Preventive Nutrition and Food Science
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• 제6권4호
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• pp.230-234
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• 2001

The extract of Ulmus davidiana var. japonica cortex has known as natural anti-inflammatory substance in East Asia. For the identification of antimicrobial substance, it was extracted by using methanol and fractionated by using different organic solvents. The fraction of butanol was represented the highest antimicrobial activities. Therefore, the butanol fraction was purified and identified the chemical structure by $^1$H and $^{13}$ C-NMR spectra, FT-IR and EI/MS spectroscopies. The isolated antimicrobial substance was identified as cis-2-[3,4-dihydroxy phenyl]-3,4-dihydro-2H-1-benzopyran-3,5,7-triol, which has commonly known as (-)epicatechin. Its minimum inhibitory concentrations (MICs) against Staphylococcus aureus and Listeria monocytogenes were shown as 100 $\mu\textrm{g}$/mL, respectively.