• Title/Summary/Keyword: Bacillus broth

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Isolation and Antimicrobial Action of Growth Inhibitory Substance on Food-borne Microorganisms from Dryopteris crassirhizoma Nakai (관중(Dryopteris crassirhizoma Nakai)의 식중독 미생물 증식 억제 물질의 분리 및 항균작용)

  • Han, Ji-Sook;Lee, Ji-Young;Baek, Nam-In;Shin, Dong-Hwa
    • Korean Journal of Food Science and Technology
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    • v.33 no.5
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    • pp.611-618
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    • 2001
  • The ethanol extract of Dryopteris crassirhizoma Nakai showed strong growth inhibition against 5 strains of Listeria monocytogenes at the concentrations of $100{\sim}500$ ppm and the minimum inhibitory concentration of n-hexane fraction was under 50 ppm. The D8-2-5 fraction isolated from n-hexane fraction of Dryopteris crassirhizoma Nakai showed a strong bactericidal activity on 5 strains of L. monocytogenes at 20 ppm level in tryptic soy broth medium. At the level, the viable count was reduced $4{\sim}6$ log cycle compared to initial cell number. Observation by the measurement of adenosine triphosphate (ATP) contents and transmission electron microscope showed that disruptions of the cell wall and elution of intracellular ATP are assumed to be due to the bactericidal activity. In addition, the n-hexane fraction of Dryopteris crassirhizoma Nakai showed the strong growth inhibitions at 50 ppm on Vibrio parahaemolyticus and Bacillus cereus, and at 25 ppm on Staphylococcus aureus.

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Cold-Sensitive Growth of Bacillus subtilis Mutants Deleted for Putative DEAD-Box RNA Helicase Genes (Bacillus subtilis DEAD-Box RNA Helicase 유전자 결손 균주들의 저온 민감성 생장)

  • Oh, Eun-Ha;Lee, Sang-Soo
    • Korean Journal of Microbiology
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    • v.46 no.3
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    • pp.233-239
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    • 2010
  • Four genes (yqfR, yfmL, ydbR, deaD) were identified as putative DEAD-box RNA helicase genes in the genomic sequence of Bacillus subtilis by homology search. To understand the function of these genes, each of the genes was deleted and the constructed strains were tested for their growth charateristics at different temperatures. The growth rate of ydbR deletion mutant ($T_d$=53 min) was a little bit reduced at $37^{\circ}C$ as compared to that of wild type strain (CU1065). But the growth rate of other three (yqfR, yfmL, deaD) deletion mutants ($T_d$=30-40 min) is nearly equal to the growth rate of wild type ($T_d$=32 min). On the other hands, the growth rate of deletion mutants were reduced at $22^{\circ}C$ in order of yqfR ($T_d$=151 min), yfmL ($T_d$=214 min), ydbR ($T_d$=343 min), which showed cold-sensitive phenotype. The deletion mutant of deaD ($T_d$=109 min) grew equally as compared to the growth rate ($T_d$=102 min) of the wild type at $22^{\circ}C$ and did not show cold-sensitive growth. Double, triple and quadruple deletion mutants of these genes were constructed, and growth rate of these mutants were measured at various temperature conditions ($22^{\circ}C$, $37^{\circ}C$, $42^{\circ}C$) using LB broth. Multiple deletion mutations showed more severe cold-sensitive growth than single deletion mutations, and double deletion of ydbR and yfmL ($T_d$=984 min) showed most cold-sensitive growth than any other double mutants. Such a cold-sensitive growth of these mutations is quite similar to the result of csdA or srmB deletion in E. coli and suggested that physiological role of ydbR and yfmL is related with ribosome assembly.

Comparison of DNA Extraction Methods for the Detection of Foodborne Pathogenic Bacteria from Livestock Manure Composts (퇴비에서 식중독균 검출을 위한 DNA 추출 방법 비교)

  • Kim, Sung-Youn;Seo, Dong-Yeon;Moon, Ji-Young
    • Journal of Food Hygiene and Safety
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    • v.34 no.6
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    • pp.557-561
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    • 2019
  • This study investigated the efficacy of DNA extraction methods for real-time PCR detection of foodborne pathogenic bacteria in livestock manure composts. Livestock manure composts were inoculated with Escherichia coli O157:H7, Salmonella Typhimurium, Listeria monocytogenes, Bacillus cereus and incubated in enrichment broth. For DNA extraction, enriched samples were treated following boiling method, by chloroform, C18 powder, and proteinase K. As a result, 4 species of bacteria were detected by real-time PCR when subjected to boiling for 30 min and treated with proteinase K. These results suggest that detection of foodborne pathogens by real-time PCR from livestock manure composts could be applicable using effective DNA extraction methodology such as the boiling method or proteinase K.

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.

Isolation and In vitro and In vivo Antifungal Activity of Phenylacetic acid Produced by Micromonospora aurantiaca Strain JK-1

  • Kim, Hyo-Jin;Hwang, In-Sun;Kim, Beom-Seok;Hwang, Byung-Kook
    • The Plant Pathology Journal
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    • v.22 no.1
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    • pp.75-89
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    • 2006
  • The actinomycete strain JK-1 that showed strong inhibitory activity against some plant pathogenic fungi and oomycetes was isolated from Jung-bal Mountain in Ko-yang, Korea. The strain JK-1 produced spores singly borne on sporophores and the spores were spherical and 0.9-1.2 11m in diameter. The cell wall of the strain JK-1 contained meso-diaminopimelic acid. The actinomycete strain JK-1 was identified as the genus Micromonospora based on the morphological, physiological, biochemical and chemotaxonomic characteristics. From the 168 rDNA analysis, the strain JK-1 was assigned to M aurantiaca. The antibiotic MA-1 was purified from the culture broth of M aurantiaca JK-1 using various purification procedures, such as Diaion HP20 chromatography, C18 flash column chromatography, silica gel flash column chromatography and Sephadex LH-20 column chromatography. $^{1}H-$, $^{13}C-NMR$ and EI mass spectral analysis of the antibiotic MA-1 revealed that the antibiotic MA-1 is identical to phenylacetic acid. Phenylacetic acid showed in vitro inhibitory effects against fungal and oomycete pathogens Alternaria mali, Botrytis cinerea, Magnaporthe grisea, Phytophthora capsici and yeast Saccharomyces cerevisiae at < 100 $\mug$ $ml^{-1}$. In addition, phenylacetic, acid completely inhibited the growth of Sclerotinia sclerotiorum, Bacillus subtilis, Candida albicans, Xanthomonas campestris pv. vesicatoria at < $\mug$ $ml^{-1}$. Phenylacetic acid strongly inhibited conidial germination and hyphal growth of M grisea and C. orbiculare. Phenylacetic acid showed significantly high levels of inhibitory' effect against rice blast and cucumber anthracnose diseases at 250 $\mug$ $ml^{-1}$. The control efficacies of phenylacetic acid against the two diseases were similar to those of commercial compounds tricyclazole, iprobenfos and chlorothalonil .n the greenhouse.

Degradation of Lignocelluloses in Rice Straw by BMC-9, a Composite Microbial System

  • Zhao, Hongyan;Yu, Hairu;Yuan, Xufeng;Piao, Renzhe;Li, Hulin;Wang, Xiaofen;Cui, Zongjun
    • Journal of Microbiology and Biotechnology
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    • v.24 no.5
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    • pp.585-591
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    • 2014
  • To evaluate the potential utility of pretreatment of raw biomass with a complex microbial system, we investigated the degradation of rice straw by BMC-9, a lignocellulose decomposition strain obtained from a biogas slurry compost environment. The degradation characteristics and corresponding changes in the bacterial community were assessed. The results showed that rapid degradation occurred from day 0 to day 9, with a peak total biomass bacterium concentration of $3.3{\times}10^8$ copies/ml on day 1. The pH of the fermentation broth declined initially and then increased, and the mass of rice straw decreased steadily. The highest concentrations of volatile fatty acid contents (0.291 mg/l lactic acid, 0.31 mg/l formic acid, 1.93 mg/l acetic acid, and 0.73 mg/l propionic acid) as well as the highest xylanse activity (1.79 U/ml) and carboxymethyl cellulase activity (0.37 U/ml) occurred on day 9. The greatest diversity among the microbial community also occurred on day 9, with the presence of bacteria belonging to Clostridium sp., Bacillus sp., and Geobacillus sp. Together, our results indicate that BMC-9 has a strong ability to rapidly degrade the lignocelluloses of rice straw under relatively inexpensive conditions, and the optimum fermentation time is 9 days.

Identification of an Entomopathogenic Bacterium, Serratia sp. ANU101, and Its Hemolytic Activity

  • Kim, Yong-Gyun;Kim, Keun-Seob;Seo, Ji-Ae;Shrestha, Sony;Kim, Hosanna-H.;Nalini, Madanagopal;Yi, Young-Keun
    • Journal of Microbiology and Biotechnology
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    • v.19 no.3
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    • pp.314-322
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    • 2009
  • Four different bacterial colonies were isolated from an old stock of an entomopathogenic nematode, Steinernema monticolum. They all showed entomopathogenicity to final instar larvae of beet armyworm, Spodoptera exigua, by hemocoelic injection. However, they varied in colony form, susceptibility to antibiotics, and postmortem change of the infected host insects. Biolog microbial identification and 16S rDNA sequence analyses indicate that these are four different species classified into different bacterial genera. Owing to high entomopathogenicity and a cadaver color of infected insect host, Serratia sp. was selected as a main symbiotic bacterial species and analyzed for its pathogenicity. Although no virulence of Serratia sp. was detected at oral administration, the bacteria gave significant synergistic pathogenicity to fifth instar S. exigua when it was treated along with a spore-forming entomopathogenic bacterium, Bacillus thuringiensis. The synergistic effect was explained by an immunosuppressive effect of Serratia sp. by its high cytotoxic effect on hemocytes of S. exigua, because Serratia sp. caused septicemia of S. exigua when the bacterial cells were injected into S. exigua hemocoel. The cytotoxic factor(s) was present in the culture medium because the sterilized culture broth possessed high potency in the cytotoxicity, which was specific to granular cells and plasmatocytes, two main immune-associated hemocytes in insects.

A Multiplex PCR Assay for the Detection of Food-borne Pathogens in Meat Products

  • Kim, Hyoun-Wook;Kim, Ji-Hyun;Rhim, Seong-Ryul;Lee, Kyung-A;Kim, Cheon-Jei;Paik, Hyun-Dong
    • Food Science of Animal Resources
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    • v.30 no.4
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    • pp.590-596
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    • 2010
  • Meat and meat products are a potential source of food-borne pathogens, including Staphylococcus aureus, Salmonella spp., Escherichia coli O157:H7, and Bacillus cereus. A sensitive and specific PCR assay for the detection of these pathogens in meat and meat products was developed in this study, as part of a broader effort to reduce the potential health hazards posed by these pathogens. Initially, PCR conditions were standardized with purified DNA. Under standard conditions, the detection level for PCR was as low as 10 pg of purified bacterial DNA. After overnight growth of bacteria in a broth medium, as few as $10^2$ CFU of bacteria were detected by PCR assay. The primers employed in the PCR assay were found to be highly specific for individual organisms, and evidenced no cross-reactivity with heterologous organisms. Additionally, the multiplex PCR assays also amplified some target genes from the four pathogens, and multiplex amplification was obtained from as little as 10 pg of DNA, thus illustrating the excellent specificity and high sensitivity of the assay. In conclusion, this PCR-based technique provides a sensitive and specific method for the detection of S. aureus, Salmonella spp., E. coli O157:H7, and B. cereus in meat and meat products.

Solumycin : A water-soluble antifungal antibiotic from Streptomyces sp. LAM-593 (Streptomyces sp. LAM-593이 생산하는 수용성 항진균성 항생물질)

  • Yi, Dong-Heui;Park, Seung-Lim;Kwon, Tae-Jong;Chung, Ho-Kwon
    • Applied Biological Chemistry
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    • v.34 no.2
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    • pp.180-186
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    • 1991
  • A water soluble antifungal antibiotic, Solumycin, was separated from the culture broth of Streptomyces sp. LAM-593, isolated from soil, by butanol extraction, alumina-, 1st and 2nd Sephadex LH-20 column chromatography. The substance was pale yellow crystal which gave a single spot at Rf value 0.24 with ethanol-ammonia water-water (8:1:1), 0.46 with butanol-ethanol-water (5:1:4), 0.84 with 50% methanol on silica gel TLC. It was dissolved well in water, methanol and acidic aq. butanol but not in ethanol, acetone, ethyl acetate, chloroform. acetic acid etc., and gave positive Fehling and Molish reaction. The UV spectrum in methanol showed absorption at 342, 361, 380, and 404 nm. The antibiotic was active against fungi such as Candide, Cryptococcus, Saccharomyces, Trichophyton and Trichosporon, but not to bacteria such as Bacillus, Escherichia and Staphylococcus.

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Evaluation of Antioxidant, Cytoprotective and Antimicrobial Activities of the Extract and Fractions Obtained from Young Shoots of Nypa Fruticans Wurmb (니파야자(Nypa fruticans Wurmb) 싹 추출물 및 분획물의 항산화, 세포 보호 및 항균 효과에 관한 평가)

  • Shin, Hyuk Soo;Lee, Yoon Joo;Kim, Ji Woong;Song, Ba Reum;Lee, Sang Lae;Park, Soo Nam
    • Korean Journal of Pharmacognosy
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    • v.49 no.2
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    • pp.155-164
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    • 2018
  • Nypa fruticans Wurmb is a species of palm, which is widely distributed in the mangrove forest of Southeast Asia. Various parts of N. fruticans has been used as a traditional medicinal plant. However, the physiological activities of N. fruticans has not yet been clarified well. Therefore, in this study, the 50% ethanol extract and its aqueous and ethyl acetate fractions of young shoots of N. fruticans were investigated for their antioxidant, cytoprotective effect, and antimicrobial activities. Every sample possessed very high free radical and various ROS scavenging capacities assessed by employing different in vitro assays such as $DPPH^{\cdot}$, $O_2^{{\cdot}-}$, ${\cdot}OH$, and $^1O_2$ scavenging activities. Based on these results, the cytoprotective effect was investigated using the oxidative hemolysis of erythrocyte. We found that the extract and fractions provide a greater protective effect compared with (+)-${\alpha}$-tocopherol. Furthermore, antimicrobial activities were confirmed against skin pathogens by broth microdilution assay. The ethyl acetate fraction had much higher antimicrobial activities than methyl paraben against Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa, and Candida albicans. Taken together, our results indicated that the young shoots of N. fruticans may have the potential role as a natural active ingredient through their antioxidant, cytoprotective effect, and antimicrobial activities.