• Title, Summary, Keyword: bioautography

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Antimicrobial Activity and Antioxidative Activity in the Extractives of Quercus dentata Thunberg (떡갈나무 추출물의 항균활성 및 항산화활성)

  • Kim, Min-Young;Kim, Yun-Geun;Kim, Tae-Hong;Jo, Jong-Soo;Yang, Jae-Kyung
    • Journal of the Korean Wood Science and Technology
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    • v.28 no.3
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    • pp.42-51
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    • 2000
  • Antimicrobial activity and antioxidative activity of the organosoluble extractives from the leaves, bark and xylem of Quercus dentata were investigated. Antimicrobial activity was tested by paper disk method and bioautography methods, The most pronounced antimicrobial activities of leaves parts were ethyl acetate solubles fraction against the Klebsiella pneumoniae by the paper disk method. The strongest activities of bark parts were ethyl acetate solubles fraction against Bacillus subtilis, Klebsiella pneumoniae. Ethanol extractives from xylem parts showed high activities against Bacillus subtilis, Staphylococcus aureus, Klebsiella pneumoniae and Vibrio parahaemolyticus. These antimicrobial activities of ethanol extractives from Quercus dentata were in order to xylem > bark > leaves. The strong inhibition zones of the ethanol extractives and its fractions of xylem showed Rf values in 0.41~0.63. In leaves extractives, the petroleum ether solubles showed lower antioxidative activity and ethyl acetate insolubles showed higher antioxidative activity of 70% compared with the $EC_{50}$ values of the control. Antioxidative activity of bark and xylem extractives showed higher approximately 2 times than the control except the petroleum ether solubles.

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Antibacterial and Antibiofilm Activities of Diospyros malabarica Stem Extract against Streptococcus mutans (Streptococcus mutans에 대한 인도감나무 줄기 추출물의 항균활성 및 생물막 형성 억제 효과)

  • Kim, Hye Soo;Lee, Sang Woo;Sydara, Kongmany;Cho, Soo Jeong
    • Journal of Life Science
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    • v.29 no.1
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    • pp.90-96
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    • 2019
  • The objective of this study was to evaluate the potential of Diospyros malabarica stem extract, a natural materials, in oral health material. With this aim in mind, thin layer chromatography (TLC), TLC-bioautography, high-performance liquid chromatography (HPLC), electrospray ionization-mass spectrometry (ESI-MS), scanning electron microscopy (SEM), and real-time qPCR were performed. The antibacterial activity of D. malabarica stem extract against Streptococcus mutans KCTC3065 was confirmed in an n-hexane fraction with low polarity. The molecular weight of the antibacterial compound was estimated to be 188 by ESI-MS analysis. The inhibitory effects of the extract on biofilm formation and gene expression related to biofilm formation of S. mutans were determined by SEM and real-time PCR analysis. The extract inhibited the formation of S. mutans biofilms at D. malabarica stem extract concentrations of 1 mg/ml, as shown by SEM. The real-time PCR analysis showed that the expression of the gtfC gene, which is associated with biofilm formation, was significantly decreased in a dose-dependent manner. Based on the above results, it can be concluded that D. malabarica stem extracts, a natural materials, can be used in oral health products to suppress the formation of biofilms by inhibiting tooth adhesion of S. mutans, a causative agent of dental caries.

Natural Antibiotic Activity of Lactobacillus helveticus (Lactobacillus helveticus의 자연항생작용에 관한 연구)

  • Choi, Sang-Hun;Kim, Dong-Sin
    • Journal of Dairy Science and Biotechnology
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    • v.17 no.1
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    • pp.11-15
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    • 1999
  • The objective of this study was to extract and purity the antibacterial agent from the fermented milk with Lactobacillus helveticus CH-1. The extraction and purification of antibacterial agent from the Lb. helveticus fermented milk were carried out by methanol extraction, acetone extraction, Sephadex G-200 gel filteration and thin layer chromatography and the results were as followings. The antibacterial activity of methanol-acetone extraction showed antibacterial activity against test organisms, B. subtilis, E. coli, Pseu. fluorescens, Sal. typhimurium, Shi. flexneri, and Sta. aureus. Sephadex G-200 gel chromatography showed only antibacterial activity from 33 to 37th fractions of 60 fractions. The agent purified from TLC plate confirmed the antibacterial activity by the means of bioautography.

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Purification and Characterization of Natural Antifungal Protein from Astragal Seeds (Astragalus membranaceus L.). (황기 종자의 천연 항진균성 단백질의 분리정제 및 특성검정)

  • 구본성;류진창;정태영;김교창
    • Microbiology and Biotechnology Letters
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    • v.26 no.5
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    • pp.379-386
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    • 1998
  • Deterioration of food is in general caused by the presence of microorganisms and chemical compounds of food itself. There exists antimicrobial compound in the food, however, addition of food antiseptics, additives, or physico-chemical processing is a common practice. The safety of artificial chemical antiseptics became a serious public concern, therefore, new natural antiseptic compounds are in need to be developed. We have isolated a new natural antifungal protein (KBS-B2) from Astragal seed through ammonium sulfate precipitation and column chromatography using FPLC Mono-S and Superose 12HR. The purified protein inhibited growth of Candida albicans, and spore germination of food spoiling fungi such as Aspergillus ochraceus, Penicillium expensum, P. digitatum and Botrytis cineria. Antifungal effect of the KBS-B2 protein could be directly assayed by bioautography overlaying the fungal spores on the electrophoresed acrylamide gel. The comparison of N-terminal amino acid sequences of the KBS-B2 with known antifungal protein revealed that had 50% homology to thaumatin and zeamatin like proteins.

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Characterization of antimicrobial proteins produced by Bacillus sp. N32 (Bacillus sp. N32 균주가 생산하는 항균 단백질 특성)

  • Lee, Mi-Hye;Park, In-Cheol;Yeo, Yun-Soo;Kim, Soo-Jin;Yoon, Sang-Hong;Lee, Suk-Chan;Chung, Tae-Young;Koo, Bon-Sung
    • The Korean Journal of Pesticide Science
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    • v.10 no.1
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    • pp.56-65
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    • 2006
  • An antagonistic bacterial isolate, that inhibits the growth of plant pathogens, was selected and identified from 5,000 isolates screened from the rhizosphere of various crop plants. An isolate Bacillus sp. N32, tested against Colletotrichum gloeosporioides causing anthracnose disease in hot pepper, produced both a heat resistant antifungal protein and a heat sensitive antifungal protein. The heat resistant protein was partially purified by Ammonium sulfate fractionation and gel filtration chromatography. The bioautography showed that the proteins possessed high antifungal activity. The biosynthetic gene cluster responsible for the heat resistant antifungal protein was cloned from cosmid library using DNA probe obtained from PCR product with the primers targeting the conserved nucleotide sequence of the synthetic genes reported earlier, Most of the clones obtained showed higher homology to fengycin antibiotic synthetic gene family reported earlier. On the other hand, the heat sensitive protein was isolated from SDS-PAGE and electroblotting to determine the N-terminal amino acid sequences. The heat sensitive antifungal protein gene was cloned from the ${\lambda}-ZAP$ libraries using a DNA probe based on the N-terminal amino acid sequences of the heat sensitive protein. We are contemplating to clone and sequence the whole gene cluster encoding the heat sensitive protein for further analysis.

Effect of culture pH and media composition on the ratio of tcicoplanin $A_1$ and $A_2$ biosynthesis

  • Kim, Yun-Jeong;Song, Yun-Seok;No, Yong-Taek
    • 한국생물공학회:학술대회논문집
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    • pp.325-328
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    • 2001
  • Teicoplanin is a glycopeptide antibiotic produced by Actinoplanes teichomyceticus novo sp. A TCC 31121. It is active against Gram-positive bacteria and it is under evaluation for use in man. Teicoplanin mixture in fermentation broth contains major amounts of teicoplanin $A_1$ and $A_2$ and a minor amount tcicoplanin of $A_3$. Commercial teicoplanin product is composed of five major components of very similar polarity, designated T-$A_2$-l, 2, 3, 4 and 5, and the more polor component, designated T -$A_3$. The culture conditions were studied in order that hydrophilic teicoplanin $A_2$ components are more produced but hydrophobic teicoplanin $A_1$ with lower bioactivity are less produced in submerged culture. Effects of culture pH and nutrients on the biosynthes ratio of teicoplanin $A_1$ and $A_2$ were confirmed in flask culture using MOPS buffer system through TLC, bioautography and bioassay. It was elucidated that optimal pH is 7.4 for teicoplanin $A_2$ biosynthesis but is 5.2 for teicoplanin $A_1$ biosynthesis, and that trace elements stimulate $A_2$ production but malt extract stimulate $A_1$production.

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Antimicrobial Activity of Various Parts of Tomato Plants Varied with Different Solvent Extracts

  • Kim, Dong Sub;Kwack, Yurina;Lee, Jung Heon;Chun, Changhoo
    • The Plant Pathology Journal
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    • v.35 no.2
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    • pp.149-155
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    • 2019
  • The antimicrobial activity of acetone, hexane, dichloromethane, and methanol extracts from leaves, stems, immature green fruits, and red fruits of tomato plants was examined against six phytopathogens. The minimum inhibitory concentration (MIC) of the acetonic extracts from these four plant parts was lower than that of the other solvents. Among the acetonic extracts, tomato leaves had a lower MIC than the other tomato parts. The acetonic extract from tomato leaves was therefore selected as a source of antimicrobial substances. The acetonic extract from tomato leaves inhibited mycelial growth of Fusarium oxysporum f. sp. lycopersici, Glomerella cingulata, and Rhizoctonia solani. Mycelial growth of R. solani treated with acetone extract from leaves showed more susceptibility than the other phytopathogens. Using 0.31 mg/ml of the acetonic extract from leaves, mycelial growth of R. solani on days 1, 2, and 3 decreased by 50.0, 52.1, and 64.0%, respectively, compared with acetone solvent treatment. The antimicrobial compounds effective against R. solani were identified as linolenic acid and caffeic acid by bioautography and GC-MS. These two compounds were used to treat six phytopathogens to confirm their antimicrobial activities. Linolenic acid inhibited mycelial growth of R. solani, while caffeic acid showed only slight antimicrobial activity. Results indicated that we propose extracts from tomato leaves which included antimicrobial compounds may provide a new lead in the pursuit of new biological sources of agrochemical candidates.

Studies on the Antimicrobial Activities of the Extractives from Magnolia (Magnolia kobus DC. var. borealis Sarg.) (목련(Magnolia kobus DC. var. borealis Sarg.) 추출성분의 항균성에 관한 연구)

  • Kim, Yun-Geun
    • Journal of the Korean Wood Science and Technology
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    • v.27 no.1
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    • pp.105-114
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    • 1999
  • Antimicrobial activities of the organosoluble extracts, seperated fractions and isolated lignans from the leaves tissue of Magnolia kobus DC. var. borealis Sarg. were investigated. The results are summarized as follows : 1. The inhibitory components against the spore growing test were concentrated on light petroleum ether and diethyl ether soluble fractions. The light petroleum ether solubles of the leaves tissue had terpenes compound, so, that they caused growing inhibition. These appearance showed high values of Rf on TLC bioautography and GC analyses with monoterpenes. 2. In the lignans, syringaresinol(X III), medioresinol(VI), phillygenin(VIII), kobusinol A(X) showed relatively high inhibitory effects in the spore growing test, and these are all showed structural characteristic of the phenolic hydroxyl group of guaiacyl and syringyl skelecton. 3. The light petroleum ether soluble fraction showed the strongest inhibitory effect against the antimicrobial activity in the seperated fractions. 4. The inhibitory effects of the lignans against the bacteria showed not so pronounced independantly, but the extracts and separated fractions contained with these lignans showed something synergism.

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Studies on antimicrobial agent produced by lactobacillus acidophilus (Lactobacillus acidophilus가 생산한 항균물질(抗菌物質)에 관한 연구(硏究))

  • Kim, Dong-shin
    • Korean Journal of Veterinary Research
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    • v.24 no.2
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    • pp.149-162
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    • 1984
  • The research was conducted(1) to confirm the agent(s) responsible for the antimicrobial activity contained in the fermented tomato juice with L. acidophilus(2) to extract and purify the antimicrobial agent(s)(3) to find the biological, physical and chemical properties of the agent(s). The following results were obtained and summarized as followings; 1. The agent responsible for the inhibitory activity was confirmed by both well assay method using fermented tomato juice with L. acidophilus and turbidimetric technique using the cell-free filtrate or neutralized filtrate of tomato acidohilus culture and found exerted antimicrobial agent other than lactic acid. 2. The procedures of purification : The isolation and purification of antimicrobial agent from the lyophilized acidophilus tomato culture were carried out by (1) methanol extraction (2) acetone extraction, (3) Sephadex G-50 gel filtration (4) paper chromatography and (5) thin layer chromatography. 3. The biological, physical and chemical properties of antimicrobial agent: The biological, physical, chemical properties of the purified antimicrobial agent were: (1) The antimicrobial activity was strong against test organisms; Bacillus subtilis(ATCC 6633), Escheichia coli(ATCC 25922), Staphylococcus aureus(ATCC 167), Pseudomonas fluorescens(KFCC 32394), Proteus vulgaris and Shigella dysenteriae. (2) The pH value of the agent was 2.0 and the inhibitory activity was lost when it was neutralized at 7.0 of pH and the agent was heat stable at $121^{\circ}C$ for 60 minutes. (3) The ultraviolet light absorption spectra of methanol-acetone extract and TLC fraction exhibited a maximum absorption at 260nm and 224nm respectively. (4) The most purified agent from TLC plate increased about 130-fold in activity. (5) The agent isolated from TLC plate was free from $H_2O_2$ or lactic acid. 4. Bioautographic assy: By means of bioautography of the agent on silica gel of TLC plate a strong inhibitory activity against B. subtilis was demonstrated. 5. Mass spectrometry: The agent obtained from TLC plate was analyzed by mass spectrometry which show the parent peak at m/e 264 suggesting the molecular weight of the compound and molecular group such as [$C_2H{^+}_4$], [CO], [CH=NH], [$C_3{H^}4_7$], [$\begin{array}{rcl}O\\{\parallel}\\CH_3-C\\\end{array}$], [$C_6-H{^+}_{11}$], [$C_5H{^+}_{11}$], [$\begin{array}{rcl}O\\{\parallel}\\C_5H_7-C^+\\\end{array}$] were suggested.

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Characterization of a heat-resistant antimicrobial peptide secreted by Bacillus subtilis A405 (Bacillus subtilis A405 균주가 생성하는 내열성 항균 peptide의 특성 검정)

  • Koo, Bon-Sung;Lee, Seung-Bum;Yoon, Sang-Hong;Song, Gae-Kyung;Chung, Dae-Sung;Byun, Myung-Ok;Ryu, Jin-Chang
    • The Korean Journal of Pesticide Science
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    • v.2 no.3
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    • pp.28-35
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    • 1998
  • An antimicrobial peptide producing bacterium, Bacillus subtilis A405, was screened and identified among 700 of antagonistic bacteria. The heat-resistant antimicrobial peptide, AMP-405, was purified from the broth culture of B. subtilis A405 through $20{\sim}40%$ ammonium sulfate precipitation and ultrafiltration. The AMP-405 exhibited strong antimicrobial activities against Botrytis cinerea, Cercospora sp., Fusarium oxysporum, Penicillium digitatum, Celletotrichum gloeosporioides, Rhizoctonia solani, Pythium ultimum, Pyricularia oryzae, Escherichia coli, Pseudomonas spp. and Candida albicans. The molecular weight of the peptide was about 3.0 kDa determined by SDS-PAGE, Native-PAGE and Tris-Tricine gradient electrophoresis, and composed of 9 kinds of amino acid such as aspartic acid, glycine, serine, glutamine, valine, leucine, isoleucine, proline, tyrocine. To determine the efficiency of AMP-405 as a potential maintenance of fruits freshness, cherry tomato was srored at $25^{\circ}C$ for 2 weeks after treatment with $50{\mu}g/ml$ of AMP-405 and $10^{5}$ spores/ml of Botrytis cinerea simultaneously. Treatment with AMP-405 resulted in significantly less infection by Botrytis cinerea, than the treatment with tap water as a control.

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