• Title/Summary/Keyword: cell-free supernatant

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Anti-Biofilm Activity of Cell-Free Supernatant of Saccharomyces cerevisiae against Staphylococcus aureus

  • Kim, Yeon Jin;Yu, Hwan Hee;Park, Yeong Jin;Lee, Na-Kyoung;Paik, Hyun-Dong
    • Journal of Microbiology and Biotechnology
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    • v.30 no.12
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    • pp.1854-1861
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    • 2020
  • Staphylococcus aureus is one of the most common microorganisms and causes foodborne diseases. In particular, biofilm-forming S. aureus is more resistant to antimicrobial agents and sanitizing treatments than planktonic cells. Therefore, this study aimed to investigate the anti-biofilm effects of cell-free supernatant (CFS) of Saccharomyces cerevisiae isolated from cucumber jangajji compared to grapefruit seed extract (GSE). CFS and GSE inhibited and degraded S. aureus biofilms. The adhesion ability, auto-aggregation, and exopolysaccharide production of CFS-treated S. aureus, compared to those of the control, were significantly decreased. Moreover, biofilm-related gene expression was altered upon CFS treatment. Scanning electron microscopy images confirmed that CFS exerted anti-biofilm effects against S. aureus. Therefore, these results suggest that S. cerevisiae CFS has anti-biofilm potential against S. aureus strains.

Isolation of Bacteria Associated with the King Oyster Mushroom, Pleurotus eryngii

  • Lim, Yun-Jung;Ryu, Jae-San;Shi, Shanliang;Noh, Won;Kim, Eon-Mi;Le, Quy Yang;Lee, Hyun-Sook;Ro, Hyeon-Su
    • Mycobiology
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    • v.36 no.1
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    • pp.13-18
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    • 2008
  • Eight distinct bacteria were isolated form diseased mycelia of the edible mushroom, Pleurotus eryngii. 16S rDNA sequence analysis showed that the isolates belonged to a variety of bacterial genera including Bacillus (LBS5), Enterobacter (LBS1), Sphingomonas (LBS8 and LBS10), Staphylococcus (LBS3, LBS4 and LBS9) and Moraxella (LBS6). Among them, 4 bacterial isolates including LBS1, LBS4, LBS5, and LBS9 evidenced growth inhibitory activity on the mushroom mycelia. The inhibitory activity on the growth of the mushroom fruiting bodies was evaluated by the treatment of the bacterial culture broth or the heat-treated cell-free supernatant of the broth. The treatment of the culture broths or the cell-free supernatants of LBS4 or LBS9 completely inhibited the formation of the fruiting body, thereby suggesting that the inhibitory agent is a heat-stable compound. In the case of LBS5, only the bacterial cell-containing culture broth was capable of inhibiting the formation of the fruiting body, whereas the cell-free supernatant did not, which suggests that an inhibitory agent generated by LBS5 is a protein or a heat-labile chemical compound, potentially a fungal cell wall-degrading enzyme. The culture broth of LBS1 was not inhibitory. However, its cell-free supernatant was capable of inhibiting the formation of fruiting bodies. This indicates that LBS1 may produce an inhibitory heat-stable chemical compound which is readily degraded by its own secreted enzyme.

Concentration of cell-free fermentation broth containing poly (${\gamma}-glutamic acid$) by ultrafiltration

  • Do, Jin-Hwan;Jang, Ho-Nam;Lee, Sang-Yeop
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.45-46
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    • 2000
  • After cell removal from fermentation broth the supernatant containing poly (${\gamma}-glutamic acid$) (${\gamma}-PGA$) was concentrated by ultrafiltration in order to reduce the amount of organic solvents such as methanol, ethanol, propanol required for the recover of ${\gamma}-PGA$ with precipitation. The concentration and volume reduction of cell-free solution by ultrafiltration could reduce 3 times the amount of ethanol for the recover of ${\gamma}-PGA$ from cell-free fermentation broth.

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Inhibition of Aflatoxin Production of Aspergillus flavus by Lactobacillus casei

  • Chang, In-Jeong;Kim, Jeong-Dong
    • Mycobiology
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    • v.35 no.2
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    • pp.76-81
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    • 2007
  • Lactobacillus casei KC-324 was tested for its ability to inhibit aflatoxin production and mycelial growth of Aspergillus flavus ATCC 15517 in liquid culture. flatoxin $B_{1}$ biosynthesis and mycelial growth were inhibited in both simultaneous culture and individual antagonism assays, suggesting that the inhibitory activity was due to extracellular metabolites. produced in cell-free supernatant fluids of the cultured broth of L. casei KC-324. In cell-free supernatant fluids of all media tested, deMan, Rogosa and Sharpe broth, potato dextrose broth, and Czapek-Dox broth+1% yeast extract showed higher antiaflatoxigenic activity. In these case, fungal growths, however, was not affected as measured by mycelial dry weight. The antiaflatoxigenic metabolites from L. casei KC-324 were produced over wide range of temperatures between $25^{\circ}C$ and $37^{\circ}C$. However, these metabolites were not thermostable since the inhibitory activity of the supernatant was inactivated within 30 minutes at $100^{\circ}C$ and $121^{\circ}C$. The inhibitory activity was not influenced by changing pH of supernatant between 4 and 10. However, the antiaflatoxigenic activity was slightly reduced at pH 10.

INTRACELLULAR AMINO ACID PROFILE OF RUMEN BACTERIA AS INFLUENCED BY UREA FEEDING AND ITS DURATION

  • Kobayashi, Y.;Wakita, M.;Hoshino, S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.6 no.4
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    • pp.619-622
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    • 1993
  • Rumen bacterial amino acids in sheep on urea diet were monitored to assess a possible change in amino acid synthesis as a long term response to high rumen ammonia environment. A sheep was fed a semipurified diet with soybean meal, followed by a diet with urea as a main nitrogen source. Mixed rumen bacteria were harvested from ruminal fluid taken 3 h after feeding (twice in soybean meal feeding and 6 times in urea feeding) and fractionated as cell wall, proteins and protein-free cell supernatant of monitor amino acids in each fraction. Ruminal ammonia concentration at the sampling ranged from 5.7 to 39.5 mgN/dl. Cell wall and protein fractions of mixed rumen bacteria were stable in their amino acid composition regardless of nitrogen sources of diet and the feeding duration. However, protein-free cell supernatant fraction showed a higher alanine proportion with urea feeding (18.6 and 28.2 molar % of alanine for samples from sheep fed soybean meal and urea, respectively) and its duration (20.6 and 32.9 molar % for samples from sheep on urea diet for 1 and 65 days, respectively). Total free amino acid level of bacteria was depressed in the initial period of urea feeding but restored on 65th day of the feeding. These results suggest that an alanine synthesizing system may develop in rumen bacteria as urea feeding becomes longer.

Antibacterial and Antibiofilm Effect of Cell-Free Supernatant of Lactobacillus brevis KCCM 202399 Isolated from Korean Fermented Food against Streptococcus mutans KCTC 5458

  • Kim, Jong Ha;Jang, Hye Ji;Lee, Na-Kyoung;Paik, Hyun-Dong
    • Journal of Microbiology and Biotechnology
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    • v.32 no.1
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    • pp.56-63
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    • 2022
  • This study aims to determine the antibiofilm effect of cell-free supernatant (CFS) of Lactobacillus brevis strains against Streptococcus mutans strains. To study the antibiofilm mechanism against S. mutans strains, antibacterial effects, cell surface properties (auto-aggregation and cell surface hydrophobicity), exopolysaccharide (EPS) production, and morphological changes were examined. The antibiofilm effect of L. brevis KCCM 202399 CFS as morphological changes were evaluated by scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM), compared with the control treatment. Among the L. brevis strains, L. brevis KCCM 202399 showed the highest antibiofilm effect on S. mutans KCTC 5458. The antibacterial effect of L. brevis KCCM 202399 against S. mutans KCTC 5458 was investigated using the deferred method (16.00 mm). The minimum inhibitory concentration of L. brevis KCCM 202399 against S. mutans KCTC 5458 was 25.00%. Compared with the control treatment, L. brevis KCCM 202399 CFS inhibited the bacterial adhesion of S. mutans KCTC 5458 by decreasing auto-aggregation, cell surface hydrophobicity, and EPS production (45.91%, 40.51%, and 67.44%, respectively). L. brevis KCCM 202399 CFS inhibited and eradicated the S. mutans KCTC 5458 biofilm. Therefore, these results suggest that L. brevis KCCM 202399 CFS may be used to develop oral health in the probiotic industry.

Anti-Bacterial Effect of Lactobacillus rhamnosus Cell-Free Supernatant Possessing Lysozyme Activity Against Pathogenic Bacteria (라이소자임 활성을 보유한 Lactobacillus rhamnosus 배양물의 병원성 미생물에 대한 항균 효과)

  • Lee, Jiyeon;Lim, Hyeji;Kim, Misook
    • Journal of the Korean Dietetic Association
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    • v.24 no.4
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    • pp.330-343
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    • 2018
  • Recently, there has been a growing demand for natural preservatives because of increased consumer interest in health. In this study, we produced Lactobacillus rhamnosus cell-free supernatant (LCFS) and evaluated and compared its antimicrobial activity with existing natural preservatives against pathogenic microorganisms and in chicken breast meat contaminated with Escherichia coli and Staphylococcus aureus. Lactobacillus rhamnosus cell-free supernatant possessed 30 units of lysozyme activity and contained 18,835 mg/L of lactic acid, 2,051 mg/L of citric acid and 5,060 mg/L of acetic acid. Additionally, LCFS inhibited the growth of fourteen pathogenic bacteria, S. aureus, Bacillus cereus, Listeria monocytogenes, Vibrio parahaemolyticus, Listeria innocua, S. epidermidis, L. ivanovii, E. coli, Pseudomonas aeruginosa, Shigella sonnei, Shi. flexneri, Proteus vulgaris, Pseudomonas fluorescens, and Klebsiella pneumoniae. The antibacterial activity of LCFS was stronger than that of egg white lysozyme (EWL), Durafresh (DF) and grapefruit seed extract (GSE). Additionally, LCFS maintained its antimicrobial activity after heat treatment at $50^{\circ}C{\sim}95^{\circ}C$ and at pH values of 3~9. Moreover, LCFS inhibited the growth of E. coli and S. aureus in chicken breast meat. In conclusion, it is expected that LCFS, which contains both lysozyme and three organic acids, will be useful as a good natural preservative in the food industry.

Reduction of hexavalent chromium by pseudomonas aeruginosa HP014

  • Oh, Young-Sook;Choi, Sung-Chan
    • Journal of Microbiology
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    • v.35 no.1
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    • pp.25-29
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    • 1997
  • Microbial reduction of hexavalent (VI) to trivalent (III) chromium decreases its toxicity by two orders of magnitude. In order to investigate the nature of Cr-reduction, Cr-resistant Pseudomonas aeruginosa HP014 was isolated and tested for its reduction capability. At the concentration of 0.5 mM Cr(VI), cell growth was not inhibited by the presence of Cr(VI) in a liquid medium, and Cr(VI) reduction was accompanied by ell growth. When cell-free extract was tested, the reduction of Cr(VI) showed a saturation kinetics with the maximum specific activity of 0.33 .mu.mol min$\^$-1/ mg$\^$-1/ cell protein, and an apparent K. of 1.73 mM Cr(VI). The activity required either NADH or NADPH as an electron donor. However, NADPH gave 50% as mush activity as sequently the supernatant and pelleted membrane fractions were tested for Cr(VI) reduction activity. The supernatant of the centrifugation showed almost the same Cr(VI) reduction activity as compared with that of the cell-free extract, indicating that the Cr(VI)-reducing activity of P. aeruginosa HP-14 is due to soluble enzyme. Moreover, the activity appeared to be the highest among the known activities, suggesting that the strain might be useful for remediation of Cr(VI)-contaminated sites.

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Use of Vegetable Waste as a Culture Medium Ingredient Improves the Antimicrobial and Immunomodulatory Activities of Lactiplantibacillus plantarum WiKim0125 Isolated from Kimchi

  • Seul-Gi Jeong;Ho Myeong Kim;Moeun Lee ;Jung Eun Yang;Hae Woong Park
    • Journal of Microbiology and Biotechnology
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    • v.33 no.1
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    • pp.75-82
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    • 2023
  • Lactic acid bacteria (LAB) isolated from kimchi (a traditional Korean dish typically made of fermented cabbage) can provide various health benefits, including anti-obesity, antioxidant, antiinflammatory, anticancer, and antimicrobial effects. In this study, we examined the antimicrobial and immunomodulatory effects of Lactiplantibacillus plantarum WiKim0125 cultured in de Man, Rogosa, and Sharpe (MRS) medium containing vegetable waste. Live bacterial cells were eliminated via supernatant filtration or heat treatment. The cell-free supernatant (CFS) obtained from culture broth containing kimchi cabbage waste (KCW), cabbage waste (CW), or onion waste (OW) showed significantly higher antimicrobial activity against skin pathogens (Propionibacterium acnes and Staphylococcus aureus) and foodborne pathogens (Escherichia coli and Salmonella typhimurium), with inhibition zones ranging between 4.4 and 8.5 mm, compared to that in conventional MRS medium (4.0-7.3 mm). In lipopolysaccharide-stimulated RAW264.7 cells, both supernatant and heat-inactivated Lb. plantarum WiKim0125 from culture media containing KCW and CW suppressed the production of inflammatory cytokines (72.8% and 49.6%, respectively) and nitric oxide (62.2% and 66.7%, respectively) without affecting cell viability. These results indicate that vegetable waste can potentially increase the antimicrobial and immunoregulatory potency of LAB while presenting a molecular basis for applying postbiotics to health products.

Inhibition of the Aspergillus flavus Growth and Aflatoxin B1 Contamination on Pistachio Nut by Fengycin and Surfactin-Producing Bacillus subtilis UTBSP1

  • Farzaneh, Mohsen;Shi, Zhi-Qi;Ahmadzadeh, Masoud;Hu, Liang-Bin;Ghassempour, Alireza
    • The Plant Pathology Journal
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    • v.32 no.3
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    • pp.209-215
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    • 2016
  • In this study, the treatment of pistachio nuts by Bacillus subtilis UTBSP1, a promising isolate to degrade aflatoxin B1 (AFB1), caused to reduce the growth of Aspergillus flavus R5 and AFB1 content on pistachio nuts. Fluorescence probes revealed that the cell free supernatant fluid from UTBSP1 affects spore viability considerably. Using high-performance liquid chromatographic (HPLC) method, 10 fractions were separated and collected from methanol extract of cell free supernatant fluid. Two fractions showed inhibition zones against A. flavus. Mass spectrometric analysis of the both antifungal fractions revealed a high similarity between these anti-A. flavus compounds and cyclic-lipopeptides of surfactin, and fengycin families. Coproduction of surfactin and fengycin acted in a synergistic manner and consequently caused a strong antifungal activity against A. flavus R5. There was a positive significant correlation between the reduction of A. flavus growth and the reduction of AFB1 contamination on pistachio nut by UTBSP1. The results indicated that fengycin and surfactin-producing B. subtilis UTBSP1 can potentially reduce A. flavus growth and AFB1 content in pistachio nut.