• Title/Summary/Keyword: Activity Sequencing

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Biosurfactant Production from Novel Air Isolate NITT6L: Screening, Characterization and Optimization of Media

  • Vanavil, B.;Perumalsamy, M.;Rao, A. Seshagiri
    • Journal of Microbiology and Biotechnology
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    • v.23 no.9
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    • pp.1229-1243
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    • 2013
  • In this paper, an air isolate (NITT6L) has been screened based on hemolytic activity, emulsification activity, drop collapsing test, and oil displacement test, as well as lipase activity. It was found that strain NITT6L was able to reduce the surface tension of the medium from 61.5 to 39.83 mN/m and could form stable emulsions with tested vegetable oils. Morphological, biochemical, 16S rRNA sequencing analyses, and fatty acid methyl ester analysis using gas chromatography confirmed that the air isolate under study was Pseudomonas aeruginosa. Characterization of the biosurfactant using agar double diffusion assay revealed that the biosurfactant was anionic in nature, and CTAB-methylene blue assay and Molisch test revealed its glycolipid nature. The FT-IR spectrum confirmed that the crude biosurfactant was a rhamnolipid. Using unoptimized medium containing sucrose as the carbon source, the isolate was found to produce 0.3 mg/ml of rhamnolipid in batch cultivation (shake flask) at $37^{\circ}C$ and pH 7. Optimization of the medium components was carried out using design of experiments and the yield of rhamnolipid has been enhanced to 4.6 mg/ml in 72 h of fermentation.

Characterization and Antibacterial Activity of Lactobacillus casei HK-9 Isolated from Korean Rice Wine, Makgeolli (막걸리에서 분리한 젖산세균인 Lactobacillus casei HK-9의 특성 및 항균 활성)

  • Baek, Hyun;Choi, Moon-Seup;Oh, Kye-Heon
    • KSBB Journal
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    • v.27 no.3
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    • pp.161-166
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    • 2012
  • The purpose of this work was to examine the antibacterial activity derived from a lactic acid bacterium isolated from korean rice wine, called makgeolli. Various physiological and biochemical properties of this strain were characterized. Both the BIOLOG system and phylogenetic analysis using 16S rRNA sequencing were utilized for identification, and the strain was designated as Lactobacillus casei HK-9, and registered in GenBank as [JQ951606]. Growth rate, production of organic acids (e.g., lactic acid and acetic acid), and pH changes during growth were monitored. The maximum concentrations of lactic acid and acetic acid were approximately 576 mM and 199 mM, respectively, and pH was changed from 7.00 to 3.74 after 72 h of incubation. HPLC was used to confirm the production of lactic acid and acetic acid. Significant antimicrobial activity of the concentrated supernatant was demonstrated against various food-poison causing bacteria (e.g., Bacillus cereus, Listeria monocytogenes, Staphylococcus aureus, methicillin-resistant Staphylococcus aureus, Escherichia coli, Salmonella enteritidis). Ethanol tolerance of L. casei HK-9 showed up to 12% of ethanol within the culture.

Probiotic Properties of Lactobacillus plantarum NK181 Isolated from Jeotgal, a Korean Fermented Food

  • Lee, Na-Kyoung;Kim, Hyoun-Wook;Chang, Hyo-Ihl;Yun, Cheol-Won;Kim, Seung-Wook;Kang, Chang-Won;Paik, Hyun-Dong
    • Food Science and Biotechnology
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    • v.15 no.2
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    • pp.227-231
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    • 2006
  • Strain NK181 was isolated for probiotic use from jeotkal and based on results of API 50 CHL kit and 16S rDNA sequencing was tentatively named Lactobacillus plantarum NK181. L. plantarum NK181 was highly resistant to artificial gastric juice (pH 2.5) and bile acid and demonstrated strong adherence to Caco-2 cells. In test using API ZYM kit, eight enzymes were produced. Supernatant of L. plantarum NK181 exhibited about 30% 1,1-diphenyl-2-picyryl hedrazyl (DPPH) radical-scavenging activity and reduced cholesterol by 70%. These results demonstrate potential use of L. plantarum NK181 as health-promoting probiotic.

Screening of Bacteriocin-producing Bacillus Strains Isolated from Domestic Animal Feces for Antagonistic Activities against Clostridium perfringens

  • Han, Sun-Kyung;Choi, Hyun-Jong;Lee, Sang-Myeong;Shin, Myeong-Su;Lee, Wan-Kyu
    • Food Science of Animal Resources
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    • v.31 no.3
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    • pp.405-412
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    • 2011
  • The purpose of this study was to isolate and characterize bacteriocin-producing bacteria against Clostridium perfringens from domestic animals to determine their usefulness as probiotics. The feces of cattle and chicken were used as sources to isolate bacteriocin-producing bacteria using the spot-on-lawn method. In total, 900 bacterial stains were isolated from domestic animal feces, and 19 strains were finally selected after determining the inhibitory activity against the pathogenic indicator C. perfringens KCTC 3269. Eighteen strains of Bacillus subtilis and one strain of Brevibacillus parabrevis were identified by 16s rRNA sequencing. Most of the bacterial strains isolated were resistant to 0.5% bile salts and remained viable after 2 h at pH 3.0. Additionally, some B. subtilis strains showed strong inhibitory activity against Listeria monocytogenes. We isolated and screened B. subtilis strains CB 153 and CB 189 from cattle and B. subtilis MSC 156 and B. parabrevis MSC 164 from chickens using probiotic selection criteria such as inhibition activity against C. perfringens and tolerance to acid and bile salts. The isolated bacteriocin-producing bacteria and/or bacteriocin have the potential to be used as probiotics in the livestock industry.

Studies on the Development of a Microbial Cryoprotectant Formulation Using a W/O/W Multiple Emulsion System

  • Bae, Eun-Kyung;Cho, Young-Hee;Park, Ji-Yong
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.673-679
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    • 2004
  • A microbial cryoprotectant formulation using a W/O/W multiple emulsion system was developed. The psychrotolerant microorganism, B4, isolated from soil in South Korea, was observed by the drop freezing method, in which the microorganism sample inhibited ice nucleation activity. The antifreeze activity was eliminated when the microorganism sample was treated with protease, indicating that the antifreeze activity was due to the presence of antifreeze protein. The result of the l6S rDNA sequencing indicated the B4 strain was most closely related to a species of the genus Bacillus. Culture broth of B4 strain (Bacillus sp.) and rapeseed oil containing 1 % polyglycerine polyricinolate (PGPR) were used as core and wall material, respectively. The most stable W/O emulsion was prepared at a core/oil ratio of 1:2. The highest W/O/W emulsion stability was achieved when the primary emulsion to external aqueous phase containing 0.5% caster oil polyoxyethylene ether $(COG25^{TM})$ ratio was 1:1. Microcrystalline cellulose showed better W/O/W emulsion stability than other polymer types. The viability of cells in a W/O/W emulsion was higher than free cells during storage at $37^\circ{C}$. An acidic pH and UV exposure decreased the viability of free cells, but cells in W/O/W emulsion were more stable under these conditions.

Bioproduction and Anticancer Activity of Biosurfactant Produced by the Dematiaceous Fungus Exophiala dermatitidis SK80

  • Chiewpattanakul, Paramaporn;Phonnok, Sirinet;Durand, Alain;Marie, Emmanuelle;Thanomsub, Benjamas Wongsatayanon
    • Journal of Microbiology and Biotechnology
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    • v.20 no.12
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    • pp.1664-1671
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    • 2010
  • A new biosurfactant producer was isolated from palm-oil-contaminated soil and later identified through morphology and DNA sequencing as the yeast-like fungus Exophiala dermatitidis. Biosurfactant production was catalyzed by vegetable oil, supplemented with a basal medium. The culture conditions that provided the biosurfactant with the highest surface activity were found to be 5% palm oil with 0.08% $NH_4NO_3$, at a pH of 5.3, with shaking at 200 rpm, and a temperature of $30^{\circ}C$ for a 14-day period of incubation. The biosurfactant was purified, in accordance with surfactant properties, by solvent fractionation using silica gel column chromatography. The chemical structure of the strongest surface-active compound was elucidated through the use of NMR and mass spectroscopy, and noted to be monoolein, which then went on to demonstrate antiproliferative activity against cervical cancer (HeLa) and leukemia (U937) cell lines in a dose-dependent manner. Interestingly, no cytotoxicity was observed with normal cells even when high concentrations were used. Cell and DNA morphological changes, in both cancer cell lines, were observed to be cell shrinkage, membrane blebbling, and DNA fragmentation.

Bioprospecting of Novel and Bioactive Metabolites from Endophytic Fungi Isolated from Rubber Tree Ficus elastica Leaves

  • Ding, Zhuang;Tao, Tao;Wang, Lili;Zhao, Yanna;Huang, Huiming;Zhang, Demeng;Liu, Min;Wang, Zhengping;Han, Jun
    • Journal of Microbiology and Biotechnology
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    • v.29 no.5
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    • pp.731-738
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    • 2019
  • Endophytic fungi are an important component of plant microbiota, and have the excellent capacity for producing a broad variety of bioactive metabolites. These bioactive metabolites not only affect the survival of the host plant, but also provide valuable lead compounds for novel drug discovery. In this study, forty-two endophytic filamentous fungi were isolated from Ficus elastica leaves, and further identified as seven individual taxa by ITS-rDNA sequencing. The antimicrobial activity of these endophytic fungi was evaluated against five pathogenic microorganisms. Two strains, Fes1711 (Penicillium funiculosum) and Fes1712 (Trichoderma harzianum), displayed broad-spectrum bioactivities. Our following study emphasizes the isolation, identification and bioactivity testing of chemical metabolites produced by T. harzianum Fes1712. Two new isocoumarin derivatives (1 and 2), together with three known compounds (3-5) were isolated, and their structures were elucidated using NMR and MS. Compounds 1 and 2 exhibited inhibitory activity against Escherichia coli. Our findings reveal that endophytic fungi from the rubber tree F. elastica leaves exhibit unique characteristics and are potential producers of novel natural bioactive products.

Biocontrol of Peach Gummosis by Bacillus velezensis KTA01 and Its Antifungal Mechanism

  • Tae-An Kang;GyuDae Lee;Kihwan Kim;Dongyup Hahn;Jae-Ho Shin;Won-Chan Kim
    • Journal of Microbiology and Biotechnology
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    • v.34 no.2
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    • pp.296-305
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    • 2024
  • Peach tree gummosis is a botanical anomaly distinguished by the secretion of dark-brown gum from the shoots of peach trees, and Botryosphaeria dothidea has been identified as one of the fungal species responsible for its occurrence. In South Korea, approximately 80% of gummosis cases are linked to infections caused by B. dothidea. In this study, we isolated microbes from the soil surrounding peach trees exhibiting antifungal activity against B. dothidea. Subsequently, we identified several bacterial strains as potential candidates for a biocontrol agent. Among them, Bacillus velezensis KTA01 displayed the most robust antifungal activity and was therefore selected for further analysis. To investigate the antifungal mechanism of B. velezensis KTA01, we performed tests to assess cell wall degradation and siderophore production. Additionally, we conducted reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analysis based on whole-genome sequencing to confirm the presence of genes responsible for the biosynthesis of lipopeptide compounds, a well-known characteristic of Bacillus spp., and to compare gene expression levels. Moreover, we extracted lipopeptide compounds using methanol and subjected them to both antifungal activity testing and high-performance liquid chromatography (HPLC) analysis. The experimental findings presented in this study unequivocally demonstrate the promising potential of B. velezensis KTA01 as a biocontrol agent against B. dothidea KACC45481, the pathogen responsible for causing peach tree gummosis.

Characterization and Immunomodulation Activity of Lactobacillus sakei L2 and L8 Isolated from Chicken Cecum (닭의 맹장으로부터 분리한 Lactobacillus sakei L2와 L8의 특성 및 면역활성)

  • Sim, Insuk;Park, Keun-Tae;Lim, Young-Hee
    • Microbiology and Biotechnology Letters
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    • v.44 no.2
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    • pp.201-207
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    • 2016
  • The aim of this study was to investigate the potential of lactic acid bacteria (LAB) strains as probiotics. Two strains were isolated from healthy chicken cecum and their acid and bile tolerance, residual organic acids, antibacterial activity against pathogenic bacteria, and immunomodulation activity were measured. Identification of the isolated strains was performed using the API 50CHL system and phylogenetic analysis using 16S rDNA sequencing. The isolates were determined to be Lactobacillus sakei strains. The acid tolerance of strains L2 and L8 was high enough that 75% of the inoculum survived in pH 2 for 2 h. The bile tolerance of both strains was observed at a 1% Oxgall concentration in MRS broth. The production of organic acids (lactic acid and acetic acid) and pH changes during growth were monitored and the maximum concentrations were obtained after 48 h of incubation. Culture supernatants of the two LAB strains showed strong antibacterial activity against pathogenic bacteria. The heat-killed LAB cells also induced high levels of immune cell proliferation compared with the control, and stimulated IL-6 and TNF-α production in mouse macrophages. Therefore, L. sakei strains L2 and L8 can be considered suitable probiotic bacteria.

Optimization of Hyaluronidase Inhibition Activity from Prunus davidiana (Carriere) Franch Fruit Extract Fermented by its Isolated Bacillus subtilis Strain SPF4211

  • Kim, Won-Baek;Park, So Hae;Koo, Kyoung Yoon;Kim, Bo Ram;Kim, Minji;Lee, Heeseob
    • Journal of Microbiology and Biotechnology
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    • v.26 no.9
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    • pp.1527-1532
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    • 2016
  • Strain SPF4211, having hyaluronidase (HAase) inhibition activity, was isolated from P. davidiana (Carriere) Franch fruit (PrDF) sugar extract. The phenotypic and biochemical properties based on 16S rDNA sequencing and an API 50 CHB kit suggested that the organism was B. subtilis. To optimize the HAase inhibition activity of PrDF extract by fermentation of strain SPF4211, a central composite design (CCD) was introduced based on three variables: concentration of PrDF extract (X1: 1-5%), amount of starter culture (X2: 1-5%), and fermentation time (X3: 0-7 days). The experimental data were fitted with quadratic regression equations, and the accuracy of the equations was analyzed by ANOVA. The statistical model predicted the highest HAase inhibition activity of 37.936% under the optimal conditions of X1 = 1%, X2 = 2.53%, and X3 = 7 days. The optimized conditions were validated by observation of an actual HAase inhibition activity of 38.367% from extract of PrDF fermented by SPF4211. These results agree well with the predicted model value.