• Title/Summary/Keyword: cell-free culture broth

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Antagonistic Activity against Dirty Panicle Rice Fungal Pathogens and Plant Growth-Promoting Activity of Bacillus amyloliquefaciens BAS23

  • Saechow, Sukanya;Thammasittirong, Anon;Kittakoop, Prasat;Prachya, Surasak;Thammasittirong, Sutticha Na-Ranong
    • Journal of Microbiology and Biotechnology
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    • v.28 no.9
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    • pp.1527-1535
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    • 2018
  • Bacterial strain BAS23 was isolated from rice field soil and identified as Bacillus amyloliquefaciens. Based on dual culture method results, the bacterium BAS23 exhibited potent in vitro inhibitory activity on mycelial growth against a broad range of dirty panicle fungal pathogens of rice (Curvularia lunata, Fusarium semitectum and Helminthosporium oryzae). Cell-free culture of BAS23 displayed a significant effect on germ tube elongation and mycelial growth. The highest dry weight reduction (%) values of C. lunata, H. oryzae and F. semitectum were 92.7%, 75.7%, and 68.9%, respectively. Analysis of electrospray ionization-mass spectrometry (ESI-MS) and $^1H$ nuclear magnetic resonance (NMR) spectroscopy revealed that the lipopeptides were iturin A with a C14 side chain (C14 iturinic acid), and a C15 side chain (C15 iturinic acid), which were produced by BAS23 when it was cultured in nutrient broth (NB) for 72 h at $30^{\circ}C$. BAS23, the efficient antagonistic bacterium, also possessed in vitro multiple traits for plant growth promotion and improved rice seedling growth. The results indicated that BAS23 represents a useful option either for biocontrol or as a plant growth-promoting agent.

Screening and Characterization of Microorganisms with Fibrinolytic Activity from Fermented Foods

  • Yoon, Seon-Joo;Yu, Myeong-Ae;Sim, Gwan-Sub;Kwon, Seung-Taek;Hwang, Jae-Kwan;Shin, Jung-Kue;Yeo, In-Hyun;Pyun, Yu-Rang
    • Journal of Microbiology and Biotechnology
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    • v.12 no.4
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    • pp.649-656
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    • 2002
  • Fibrinolytic microorganisms were screened from 42 samples of Korean fermented food (7 kinds of Chungook-jang, 14 kinds of commercial Doen-Jang, 5 kinds of home-made Doen-jang, and 16 kinds of Jeot-gal), 15 samples of Japanese fermented food (5 kinds of home-made soybean paste, and 10 kinds of Natto), and 19 samples of Indonesian fermented food (Tempe) as well as starters of Meju (500 microflora from Korea, and 22 from China). Initially, 11 isolates with strong fibrinolytic activity were selected for further characterization. The fibrinolytic activity of the 11 isolates ranged from 89 to 199% of standard plasmin. Four strains, M5l from Korean fermented food (Meju), I 1-1, I 1-4, and I 5-1 from Indonesian fermented food (Tempe), were chosen based on the degree of activity and reproducibility, and identified as Staphylococcus sciuri, Citrobacter or Enterobacter, Enterococcus faecalis, and Bacillus subtilis, respectively. The first two isolates are pathogenic stains while the latter two are considered as GRAS (Generally Recognized As Safe). Fibrinolytic activity of E. faecalis, characterized and designated as BRCA-5, reached a maximum, when the producer was cultivated in Ml7 broth supplemented with 1.0% glucose for 5 h at 37$^{\circ}C$ with shaking at 180 rpm. Compared to commercial fibrinolytic enzymes, the cell-free culture supernatant of 5. faecaiis BRCA-5 showed stronger activity than plasmin and streptokinase, but similar degree of specific activity as nattokinase and urokinase, aud it also demonstrated anticoagulant and antiplatelet activity ex vivo. These features of E. faecalis make it an attractive agent as a biomaterial for health-promoting foods.

A Fibrinolytic Enzyme from Bacillus amyloliquefaciens D4-7 Isolated from Chungkook-Jang; It′s Characterization and Influence of Additives on Thermostability. (청국장으로부터 분리한 Bacillus amyloliquefaciens D4-7이 분비하는 혈전용해효소의 특성 및 열안정성에 미치는 첨가물의 효과)

  • 김상숙;이주훈;안용선;김정환;강대경
    • Microbiology and Biotechnology Letters
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    • v.31 no.3
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    • pp.271-276
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    • 2003
  • Bacillus amyloliquefaciens D4, which produces a strongly fibrinolytic enzyme, was isolated from Chungkook-Jang, a traditional Korean soybean-fermented food. B. amyloliquefaciens D4 was mutated with N-methyl-N-nitro-N-nitrosoguanidine (MNNG) to yield a series of mutants with increasing levels of fibrinolytic enzyme production. After mutation, a mutant D4-7 was obtained with fibrinolytic activity about eight times stronger than the parent strain. The fibrinolytic activity of B. amyloliquefaciens D4-7, reached a maximum, when the producer was cultivated in 2% Isolated Soy Protein (ISP) broth for 48 h at $37^{\circ}C$. Compared to commercial fibrinolytic enzymes, the cell-free culture supernatant of B. amyloliquefaciens D4-7 showed stronger activity than plasmin and streptokinase. The optimum temperature and pH were $50^{\circ}C$ and 10.0 and thermostability was increased by the addition of glycerol, glucose, and NaCl.

High-Level Expression and Secretion of Bacillus pumilus Lipase B26 in Bacillus subtilis Chungkookjang

  • Lee, Mi-Hwa;Song, Jae-Jun;Choi, Yoon-Ho;Hong, Seung-Pyo;Rha, Eu-Gene;Kim, Hyung-Kwoun;Lee, Seung-Goo;Poo, Har-Young;Lee, Sang-Chul;Seu, Young-Bae;Sung, Moon-Hee
    • Journal of Microbiology and Biotechnology
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    • v.13 no.6
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    • pp.892-896
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    • 2003
  • High-level expression of the lipase B26 gene from Bacillus pumilus was achieved using Bacillus subtilis Chungkookjang isolated from the Korean traditional fermented bean paste, Chungkookjang. For the secretory production of recombinant lipase B26 in a Bacillus host system, pLipB26 was constructed by ligating the lipase B26 gene into the recently designed Escherichia coli-Bacillus shuttle vector, pLipSM, and that was then transformed into B. subtilis Chungkookjang. Among the various vector, medium, and host combinations, B. subtilis Chungkookjang harboring the pLipB26 exhibited the highest lipase activity in PY medium, and B. subtilis Chungkookjang secreted two times more enzymes than B. subtilis DB 104 under the same condition. When B. subtilis Chungkookjang harboring the pLipB26 was cultured in a 5-1 jar-fermentor containing 21 of a PY medium, the maximum lipase activity (140 U/ml) and production yield (0.68 g/l) were obtained during the late exponential phase from a cell-free culture broth. Although B. subtilis Chungkookjang also secreted extracellular proteases at the late exponential phase, these results suggested the potential of B. subtilis Chungkookjang as a host for the secretory production of foreign proteins.

Isolation and Physiological Functionality of Yeasts from Wild Flowers in Seonyudo of Gogunsanyeoldo, Jeollabuk-do, Korea (전라북도 고군산열도의 선유도 일대 야생화들로부터 효모의 분리 및 생리기능성)

  • Hyun, Se-Hee;Han, Sang-Min;Lee, Jong-Soo
    • The Korean Journal of Mycology
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    • v.42 no.3
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    • pp.201-206
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    • 2014
  • Sixty one yeast strains of 21 species were isolated from wild flowers in Gogunsanyeoldo, including Seonyudo of Jeollabuk-do, Korea, and identified by comparison of nucleotide sequences for PCR-amplified D1/D2 region of 26S rDNA using BLAST. Among them, Cryptococcus sp. including C. aureus SY1-4 were found to be dominant, and Metschnikowia sp. including M. reukaufii SY20-1 and Rhodotnula sp. such as R. ingeniosa SY1-1 were also abundantly isolated. Some physiological functionalities of the culture broth and cell-free extracts from 61 yeast strains were determined. Supernatant from Metschnilowia reukaufii SY44-6 showed anti-gout xanthine oxidase inhibitory activity of 49.6% and whitening tyrosinase inhibitory activity of 38.4%, respectively.

Evaluation of the anti-Helicobacter pylori and cytotoxic properties of the antimicrobial substances from Lactobacillus acidophilus BK13 and Lactobacillus paracasei BK57 (Lactobacillus acidophilus BK13 and Lactobacillus paracasei BK57 균주가 생산한 항균물질의 anti-Helicobacter pylori 활성 및 위장상피세포에 대한 세포독성 평가)

  • Lim, Eun-Seo
    • Korean Journal of Microbiology
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    • v.51 no.2
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    • pp.156-168
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    • 2015
  • The objective of this study is to investigate the anti-Helicobacter pylori and anti-cancer activities of the live cells (LC), cell-free culture supernatants (CFCS), and bacteriocin solution (BS) obtained from Lactobacillus acidophilus BK13 and Lactobacillus paracasei BK57 strains. After incubation for 30 h in MRS broth, the concentration of lactic acid produced by L. paracasei BK57 ($155.9{\pm}10.2mM$) was higher than in MRS broth using L. acidophilus BK13 ($126.8{\pm}7.9mM$). Maximum bacteriocin activity (128 AU/ml) of BK13 strain was observed after 30 h of cultivation at $37^{\circ}C$, however its magnitude was significantly lower than that of BK57 strain (256 AU/ml). The LC of L. acidophilus BK13 and L. paracasei BK57 were able to inhibit the growth of H. pylori ATCC 43504 at different incubation times, depending on the initial inoculum of the LAB. These CFCS and BS obtained from BK13 and BK57 strains dramatically inhibited the growth, adhesive ability, and enzymatic activity of H. pylori. Meanwhile, the anti-cancer effect of the lactic acid from L. acidophilus BK13 and L. paracasei BK57 strains on AGS cells had significant differences with the control group. Therefore, these antagonistic substances-producing strains are potentially useful as new potential antimicrobial agents for the management and prevention of H. pylori infections.