• 제목/요약/키워드: cell-free supernatant

검색결과 101건 처리시간 0.071초

Flowers in Islands and Mountains of Korea

  • Hyun, Se-Hee;Kim, Ha-Kun;Lee, Jong-Soo
    • 한국균학회소식:학술대회논문집
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    • 한국균학회 2014년도 추계학술대회 및 정기총회
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    • pp.44-44
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    • 2014
  • Yeasts were isolated from wild flowers of some islands and mountains such as Jeju-do, Ulleungdo, Yokjido, Seonyudo and Gyejoksan, Oseosan, Beakamsan and Deogyusan in Korea and were identified by comparison of nucleotide sequences for PCR-amplified D1/D2 region of 26S rDNA or internal transcribed pacer(ITS) 1 and 2 including 5.8S rDNA using BLAST. Seventy two yeast strains of two hundred eighty nine species were isolated from wild flowers in islands and mountains, Korea. Among them, Cryptococcus species were isolated the most dominantly, and Metschnikowia reukaufii were also isolated thirty species, 10.3% of total strains. Twenty-three species including Cryptococcus aureus were overlapped between yeast strains of the islands and mountains. Some physiological functionality of the culture broth and cell-free extracts from two hundred eighty nine yeast strains were determined. The supernatant of Candida sp. 78-J-2 showed antioxidant activity of 22.5%, and supernatant of Metschnilowia reukaufii SY44-6 showed anti-gout xanthine oxidase inhibitory activity of 49.6% and whitening tyrosinase inhibitory activity of 38.4%, respectively.

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Streptomyces sp. J46의 세균성구멍병원균 Xanthomonas arboricola pv. pruni에 대한 항균 활성 (Antibacterial Activity of Streptomyces sp. J46 against Bacterial Shot Hole Disease Pathogen Xanthomonas arboricola pv. pruni)

  • 이정은;임다정;김인선
    • 한국환경농학회지
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    • 제40권1호
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    • pp.20-32
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    • 2021
  • BACKGROUND: Bacterial shot hole of stone fruits is a seriuos plant disease caused by Xanthomonas arboricola pv. pruni (Xap). Techniques to control the disease are required. In this study, microorganisms with antibacterial activity were isolated to develop as a microbial agent against the bacterial shot hole. METHODS AND RESULTS: An isolate with the strongest activity among the isolates was identified as Streptomyces avidinii based on 16S rRNA gene sequence analysis and designated Streptomyces sp. J46. J46 showed suppression of bacterial leaf spot with a control value of 90% at 10 times-diluted cell free supernatant. To investigate antibacterial metabolites produced by J46, the supernatant of J46 was extracted with organic solvents, and the extracts were subjected to chromatography works. Antibacterial metabolites were not extractable with organic solvents. Both reverse and normal phase techniques were not successful because the metabolites were extremely water soluble. The antibacterial metabolites were not volatiles but protein compounds based on hydrolysis enzyme treatment. CONCLUSION: Our study suggests that Streptomyces sp. J46 may be a potential as an microbial agent against bacterial shot hole. Further study to identify the metabolites is required in more detail.

Anti-Inflammatory Response in TNFα/IFNγ-Induced HaCaT Keratinocytes and Probiotic Properties of Lacticaseibacillus rhamnosus MG4644, Lacticaseibacillus paracasei MG4693, and Lactococcus lactis MG5474

  • Ji Yeon Lee;Jeong‐Yong Park;Yulah Jeong;Chang‐Ho Kang
    • Journal of Microbiology and Biotechnology
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    • 제33권8호
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    • pp.1039-1049
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    • 2023
  • Atopic dermatitis (AD) is a chronic inflammatory disease caused by immune dysregulation. Meanwhile, the supernatant of lactic acid bacteria (SL) was recently reported to have anti-inflammatory effects. In addition, HaCaT keratinocytes stimulated by tumor necrosis factor alpha (TNF-α) and interferon gamma (IFN-γ) are widely used for studying AD-like responses. In this study, we evaluated the anti-inflammatory effects of SL from lactic acid bacteria (LAB) on TNF-α/IFN-γ-induced HaCaT keratinocytes, and then we investigated the strains' probiotic properties. SL was noncytotoxic and regulated chemokines (macrophage-derived chemokine (MDC) and thymus and activation-regulated chemokine (TARC)) and cytokines (interleukin (IL)-4, IL-5, IL-25, and IL-33) in TNF-α/IFN-γ-induced HaCaT keratinocytes. SL from Lacticaseibacillus rhamnosus MG4644, Lacticaseibacillus paracasei MG4693, and Lactococcus lactis MG5474 decreased the phosphorylation of nuclear factor-κB (NF-κB) and mitogen-activated protein kinase (MAPK). Furthermore, the safety of the three strains was demonstrated via hemolysis, bile salt hydrolase (BSH) activity, and toxicity tests, and the stability was confirmed under simulated gastrointestinal conditions. Therefore, L. rhamnosus MG4644, L. paracasei MG4693, and Lc. lactis MG5474 have potential applications in functional food as they are stable and safe for intestinal epithelial cells and could improve atopic inflammation.

Antifungical Activity of Autochthonous Bacillus subtilis Isolated from Prosopis juliflora against Phytopathogenic Fungi

  • Abdelmoteleb, Ali;Troncoso-Rojas, Rosalba;Gonzalez-Soto, Tania;Gonzalez-Mendoza, Daniel
    • Mycobiology
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    • 제45권4호
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    • pp.385-391
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    • 2017
  • The ability of Bacillus subtilis, strain ALICA to produce three mycolytic enzymes (chitinase, ${\beta}$-1,3-glucanase, and protease), was carried out by the chemical standard methods. Bacillus subtilis ALICA was screened based on their antifungal activity in dual plate assay and cell-free culture filtrate (25%) against five different phytopathogenic fungi Alternaria alternata, Macrophomina sp., Colletotrichum gloeosporioides, Botrytis cinerea, and Sclerotium rolfesii. The B. subtilis ALICA detected positive for chitinase, ${\beta}$-1,3-glucanase and protease enzymes. Fungal growth inhibition by both strain ALICA and its cell-free culture filtrate ranged from 51.36% to 86.3% and 38.43% to 68.6%, respectively. Moreover, hyphal morphological changes like damage, broken, swelling, distortions abnormal morphology were observed. Genes expression of protease, ${\beta}$-1,3-glucanase, and lipopeptides (subtilosin and subtilisin) were confirmed their presence in the supernatant of strain ALICA. Our findings indicated that strain ALICA provided a broad spectrum of antifungal activities against various phytopathogenic fungi and may be a potential effective alternative to chemical fungicides.

Effect of Lactobacillus mucosae on In vitro Rumen Fermentation Characteristics of Dried Brewers Grain, Methane Production and Bacterial Diversity

  • Soriano, Alvin P.;Mamuad, Lovelia L.;Kim, Seon-Ho;Choi, Yeon Jae;Jeong, Chang Dae;Bae, Gui Seck;Chang, Moon Baek;Lee, Sang Suk
    • Asian-Australasian Journal of Animal Sciences
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    • 제27권11호
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    • pp.1562-1570
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    • 2014
  • The effects of Lactobacillus mucosae (L. mucosae), a potential direct fed microbial previously isolated from the rumen of Korean native goat, on the rumen fermentation profile of brewers grain were evaluated. Fermentation was conducted in serum bottles each containing 1% dry matter (DM) of the test substrate and either no L. mucosae (control), 1% 24 h broth culture of L. mucosae (T1), or 1% inoculation with the cell-free culture supernatant (T2). Each serum bottle was filled anaerobically with 100 mL of buffered rumen fluid and sealed prior to incubation for 0, 6, 12, 24, and 48 h from which fermentation parameters were monitored and the microbial diversity was evaluated. The results revealed that T1 had higher total gas production (65.00 mL) than the control (61.33 mL) and T2 (62.00 mL) (p<0.05) at 48 h. Consequently, T1 had significantly lower pH values (p<0.05) than the other groups at 48 h. Ammonia nitrogen ($NH_3$-N), individual and total volatile fatty acids (VFA) concentration and acetate:propionate ratio were higher in T1 and T2 than the control, but T1 and T2 were comparable for these parameters. Total methane ($CH_4$) production and carbon dioxide ($CO_2$) were highest in T1. The percent DM and organic matter digestibilities were comparable between all groups at all times of incubation. The total bacterial population was significantly higher in T1 (p<0.05) at 24 h, but then decreased to levels comparable to the control and T2 at 48 h. The denaturing gradient gel electrophoresis profile of the total bacterial 16s rRNA showed higher similarity between T1 and T2 at 24 h and between the control and T1 at 48 h. Overall, these results suggest that addition of L. mucosae and cell-free supernatant during the in vitro fermentation of dried brewers grain increases the VFA production, but has no effect on digestibility. The addition of L. mucosae can also increase the total bacterial population, but has no significant effect on the total microbial diversity. However, inoculation of the bacterium may increase $CH_4$ and $CO_2$ in vitro.

Inhibitory Effect of Lactobacillus plantarum K11 on the Adhesion of Escherichia coli O157 to Caco-2 Cells

  • Lim, Sung-Mee;Ahn, Dong-Hyun;Im, Dong-Soon
    • Food Science and Biotechnology
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    • 제18권2호
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    • pp.343-349
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    • 2009
  • Inhibitory effect of Escherichia coli O157 adhered to Caco-2 cells by the cells of Lactobacillus plantarum K11 and the cell-free culture supernatant (CFCS) and bacteriocin prepared from this strain was investigated. As the cell counts of viable L. plantarum K11 previously adhered to Caco-2 were increased, the rate of adhesion and adherent cell counts of E. coli O157 was lower. However, because the heated L. plantarum K11 rarely have the adhesion ability to Caco-2, the adhesion rate and adherent cell counts of E. coli O157 were high. In addition, the inhibitory effects of E. coli O157 adhesion by the CFCS and bacteriocin of L. plantarum K11 were dose-dependent manner. Therefore, the inhibition of adhesion of E. coli O157 to Caco-2 may result from the antimicrobial substances such as lactic acid and bacteriocin. Moreover the inhibitory activity of adhesion by the heated bacteriocin for 30 min at 100oC was similar to activity of non-treated bacteriocin, but the activity was disappeared by treatment with protease.

A Novel Lactobacillus casei LP1 Producing 1,4-Dihydroxy-2-Naphthoic Acid, a Bifidogenic Growth Stimulator

  • Kang, Jo-Eun;Kim, Tae-Jung;Moon, Gi-Seong
    • Preventive Nutrition and Food Science
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    • 제20권1호
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    • pp.78-81
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    • 2015
  • 1,4-Dihydroxy-2-naphthoic acid (DHNA) is a bifidogenic growth stimulator (BGS) and could be a functional food ingredient since bifidobacteria are beneficial for human health. For that reason, lactic acid bacteria producing DHNA have been screened. A lactic acid bacterium LP1 strain isolated from a natural cheese was confirmed to produce DHNA, analyzed by a HPLC method. The strain was identified as Lactobacillus casei by 16S rRNA gene sequence analysis. The cell-free supernatant of fermented whey produced by L. casei LP1 presented the BGS activity for three bifidobacterial strains such as Bifidobacterium longum subsp. infantis KCTC 3127, Bifidobacterium bifidum KCTC 3202, and Bifidobacterium breve KCTC 3220 which were human-originated. To the best of our knowledge, a L. casei strain which can produce DHNA was firstly identified in this study.

Bordetella bronchiseptica의 균체성분(菌體成分)에 관한 연구(硏究) 특(特)히 DEAE-cellulose Chromatography에 의한 분획정제(分劃精制)에 대하여 (Studies on the Constituents of Bordetella bronchiseptica. Purification and Properties of the Fractions Obtained by DEAE-cellulose Column Chromatography)

  • 강병규
    • 대한수의학회지
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    • 제13권1호
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    • pp.47-52
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    • 1973
  • An attempt was carried out to purify the cell components of Bordetella bronchiseptica by the mild procedures. From the supernatant of sonic treated colls, the K-agglutinogen, hemagglutinogen and skin necrotizing factor were separated by a successive use of DEAE-cellulose column chromatography and ammonium sulfate precipitation. The specific activity of purified K-agglutinogen was demonstrated by a gel-diffusion test and was also found to be free from other biologically active substances of B. bronchiseptica: namely hemagglutinin, skin-necrotizing factor and O-antigen.

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H-Y항체의 처리가 생쥐수정란의 발달에 미치는 영향 (Effect of H-Y Antibody on in vitro Development of Mouse Embryos)

  • 고정재;심호섭;김종배;박홍양;정길생;이경광
    • 한국가축번식학회지
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    • 제10권1호
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    • pp.42-48
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    • 1986
  • These experiments were carried out to develop new techniques identifying XX-bearing embryos prior to implantation by immunological method. Antiserum to histocompatibility-Y(H-Y) antigen was prepared in adult SD(sprague-dawley) female rat by repeated immunization of newbone testis supernatant from males of the same strain. ELISA test was used to identify the H-Y antibody of antiserum. Total 124 mouse embryos (8-cell stage) were treated with H-Y antiserum and complement in BSA free Ho, pp. and Pitt's medium and cultured under the gas phase of 5% CO2 in air at 37$^{\circ}C$ for 24 to 48 hrs. The morphological characteristics of embryos treated were observed under the phase-contrast micro scope. The results obtained in these experiments were summarized as follows: 1. Optimal Density of H-Y antibody were a, pp.ared to be 0.27-0.47 by ELISA test. 2. Of total 124 embryos treated with H-Y antiserum and complement 69(55.6%) embryos developed to blastocyst and 55(44.4%) destroyed or arrested.

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Mycobacterium fortuitum의 스테로이드 9${\alpha}$-하이드록실라제의 분리 및 부분정제 (Isolation and Partial Purification of the Steroid 9${\alpha}$-Hydroxylase from Mycobacterium fortuitum)

  • 강희경
    • 약학회지
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    • 제41권5호
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    • pp.638-646
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    • 1997
  • The steroid 9${alpha}$-hydroxylase activity has been detected in cytosol fraction, $100,00{\times}g$ supernatant of cell free extract of Mycobacterium fortuitum. The activity was not linear with protein concentration in the assay suggesting 9${alpha}$-hydroxylase is a multicomponent enzyme. The 9${alpha}$-hydroxylase system was partially purified through fractional saturation of ammonium sulfate, strong anion exchange (Mono Q) column chromatography, gel filtration (Superose 12) column chromatography, and testosterone affinity gel chromatography. Ammonium sulfate 50~60% saturated fraction of the cytosol gave 9${alpha}$-hydroxylase activity. For further purification, the half-saturated ammonium sulfate fraction was applied to Mono Q, Superose 12, or affinity gel column. The purification factors of 9${alpha}$-hydroxylase containing fraction after Mono Q, Superose 12, and affinity gel chromatography was 13, 11, and 17 respectively.

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