• Title/Summary/Keyword: Diffusion in DNA

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Antimicrobial Efficacy of Fermented Dark Vinegar from Unpolished Rice (현미 발효 흑초의 항균활성)

  • Choi, Hakjoon;Gwak, Gyeongja;Choi, Dabin;Park, Jaeyoung;Cheong, Hyeonsook
    • Microbiology and Biotechnology Letters
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    • v.43 no.2
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    • pp.97-104
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    • 2015
  • Vinegar is a widely used acidic seasoning and can be manufactured using various methods and bases, including cereals, wheat, and fruits. Most studies on vinegar have been conducted to evaluate its antioxidant activity. In the present study, fermented dark vinegar (FDV) produced from unpolished rice was examined for its antimicrobial activity, biochemical content, including the amounts of sugar, total soluble sugar, organic acid, and free amino acids, and pH and physiological activity. The antimicrobial efficiency of FDV was assessed using the paper disc-agar diffusion method. FDV exhibited strong antimicrobial activity against the pathogenic bacteria and yeast strains that were tested. In fact, the activity of FDV was shown to be higher than that of the commercial antibiotics carbenicillin (50 µg/ml) and tetracycline (50 µg/ml) against Staphylococcus aureus, Escherichia coli, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella typhimurium, Yersinia enterocolitica, and Lodderomyces elongisporus. The antioxidant activity of FDV and ascorbic acid was evaluated. Using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging method, we found that FDV has the highest activity of the antioxidants. After spreading FDV onto tryptic soy broth and yeast extract-peptone-dextrose agar media, the microbial strains were isolated and characterized through physiological and biochemical analysis. Based on 16S ribosomal DNA sequence analysis, the isolated microorganisms exhibited a close similarity to Acetobacter papayae, Acetobacter pasteurianus, and Acetobacter peroxidans.

Cellulase and Xylanase Activity of Compost-promoting Bacteria Bacillus sp. SJ21 (부숙촉진 세균 Bacillus sp. SJ21 균주의 cellulase와 xylanase 활성)

  • Shin, Pyung-Gyun;Cho, Soo-Jeong
    • Korean Journal of Soil Science and Fertilizer
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    • v.44 no.5
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    • pp.836-840
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    • 2011
  • In order to isolate thermophilic compost-promoting bacteria with high activity of cellulase and xylanase, spent mushroom substrates with sawdust were collected from mushroom cultivation farm, Jinju, Gyeongnam in Korea. Among of the isolates, one strain, designated SJ21 was selected by agar diffusion method. The strain SJ21 was identified as members of the Bacillus lincheniformis by biochemical characteristics using Bacillus ID kit and VITEK 2 system. Comparative 16S rDNA gene sequence analysis showed that strain SJ21 formed a distinct phylogenetic tree within the genus Bacillus and was most closely related to Bacillus subtilis with 16S rDNA gene sequence similarity of 99%. On the basis of its physiological properties, biochemical characteristics and phylogenetic distinctiveness, strain SJ21 was classified within the genus Bacillus, for which the name Bacillus sp. SJ21 is proposed. The cellulase and xylanase activity of Bacillus sp. SJ21 was slightly increased according to bacterial population from exponential phase to stationary phase in growth curve for Bacillus sp. SJ21.

Bacterial Hash Function Using DNA-Based XOR Logic Reveals Unexpected Behavior of the LuxR Promoter

  • Pearson, Brianna;Lau, Kin H.;Allen, Alicia;Barron, James;Cool, Robert;Davis, Kelly;DeLoache, Will;Feeney, Erin;Gordon, Andrew;Igo, John;Lewis, Aaron;Muscalino, Kristi;Parra, Madeline;Penumetcha, Pallavi;Rinker, Victoria G.;Roland, Karlesha;Zhu, Xiao;Poet, Jeffrey L.;Eckdahl, Todd T.;Heyer, Laurie J.;Campbell, A. Malcolm
    • Interdisciplinary Bio Central
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    • v.3 no.3
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    • pp.10.1-10.8
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    • 2011
  • Introduction: Hash functions are computer algorithms that protect information and secure transactions. In response to the NIST's "International Call for Hash Function", we developed a biological hash function using the computing capabilities of bacteria. We designed a DNA-based XOR logic gate that allows bacterial colonies arranged in a series on an agar plate to perform hash function calculations. Results and Discussion: In order to provide each colony with adequate time to process inputs and perform XOR logic, we designed and successfully demonstrated a system for time-delayed bacterial growth. Our system is based on the diffusion of ${\ss}$-lactamase, resulting in destruction of ampicillin. Our DNA-based XOR logic gate design is based on the op-position of two promoters. Our results showed that $P_{lux}$ and $P_{OmpC}$ functioned as expected individually, but $P_{lux}$ did not behave as expected in the XOR construct. Our data showed that, contrary to literature reports, the $P_{lux}$ promoter is bidirectional. In the absence of the 3OC6 inducer, the LuxR activator can bind to the $P_{lux}$ promoter and induce backwards transcription. Conclusion and Prospects: Our system of time delayed bacterial growth allows for the successive processing of a bacterial hash function, and is expected to have utility in other synthetic biology applications. While testing our DNA-based XOR logic gate, we uncovered a novel function of $P_{lux}$. In the absence of autoinducer 3OC6, LuxR binds to $P_{lux}$ and activates backwards transcription. This result advances basic research and has important implications for the widespread use of the $P_{lux}$ promoter.

Antimicrobial Effects against Food-Borne Pathogens and Antioxidant Activity of Rhododendron brachycarpum Extract (만병초 추출물의 식중독 유발균에 대한 항균효과 및 항산화활성)

  • Choi, Moo-Young;Rhim, Tae-Jin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.10
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    • pp.1353-1360
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    • 2011
  • This study was performed to investigate the antimicrobial effects against food-borne pathogens and antioxidant activity of Rhododendron brachycarpum ethanol-extract. The antimicrobial activity of the extract was determined using a paper disc-diffusion method, and the diameter of the clear zone was measured. The diameter of the clear zone in the presence of 10 mg of extract was maximal against Bacillus cereus among the three tested Gram-positive bacteria and against Escherichia coli O157:H7 among the five tested Gram-negative bacteria. Analysis of the minimum inhibitory concentration (MIC) showed that the extract exhibited a similar efficacy as that of sorbic acid, a well-known chemical preservative. The growth inhibitory effects of the extract at concentrations of 250, 500, 1,000, and 2,000 mg/L on food-borne pathogens were determined against Staphylococcus aureus, Listeria monocytogenes, Salmonella Typhimurium, and Escherichia coli O157:H7. Growth of the microorganisms was not affected by the extract at concentrations up to 250 mg/L, but it was significantly (p<0.05) inhibited by the extract at concentrations higher than 1,000 mg/L. The antioxidant effects of the extract were examined via measurement of DPPH radical scavenging activity, inhibition of reactive oxygen species (ROS) generation using fluorescent dichlorofluorescien (DCF) assay, and prevention of peroxyl radical- and hydroxyl radical-induced supercoiled DNA breakage. The $IC_{50}$ of the extract for DPPH radical scavenging activity was about half that of ${\alpha}$-tocopherol, which was used as a positive control. DCF fluorescence intensity decreased as the concentration of the extract increased, demonstrating that ROS generation was inhibited in a concentration-dependent manner. The ROS inhibitory effect of the extract was higher than that of ascorbic acid. The extract prevented supercoiled DNA strand breakage induced by peroxyl radical and hydroxyl radical. Thus, the results of the present study demonstrate that the extract exhibits antimicrobial effects against food-borne pathogens as well as potent antioxidant capacity, suggesting that R. brachycarpum could be used as a natural antibacterial agent and effective antioxidant in food.

Isolation and Characterization of Thermophilic Bacillus licheniformis YJ09 from Spent Mushroom (Pleurotus ostreatus) Substrates (느타리 수확후배지로부터 분리된 고온성 Bacillus licheniformis YJ09의 특성)

  • Kim, Hye Soo;Kim, Chul Hwan;Cho, Soo Jeong
    • Journal of Mushroom
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    • v.14 no.4
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    • pp.244-248
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    • 2016
  • In order to isolate thermophilic bacteria with high activity of CMCase and xylanase, spent mushroom substrates was collected from an oyster mushroom cultivation farm in Jinju, Gyeongnam, Korea. Among the isolates, one strain designated as YJ09 was selected by agar diffusion method. The isolate YJ09 was identified as a member of Bacillus licheniformis based on biochemical characteristics using Bacillus ID kit and MicroLog system. Comparative 16S rDNA sequence analysis showed that isolate YJ09 formed a distinct phylogenetic tree within the genus Bacillus and was most closely related to Bacillus licheniformis with sequence similarity of 98.9%. Based on its physiological properties, biochemical characteristics and phylogenetic distinctiveness, the isolate YJ09 was classified as Bacillus licheniformis. The CMCase and xylanase activity of B. licheniformis YJ09 was slightly increased corresponding to the bacterial population from exponential phase to stationary phase in the growth curve of B. licheniformis YJ09.

Studies on Biological Activities of the Polysaccharides and Oligosaccharides of Orostachys japonicus (와송 다당체 및 올리고당류의 생리 활성)

  • Kim Ki Hoon;Kim Eun Young;Kim Yea Oon;Baek Geum Ok;Kim Han Bok;Lee Dong Seok
    • Korean Journal of Microbiology
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    • v.40 no.4
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    • pp.334-341
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    • 2004
  • Polysaccharides were prepared from Orostachys japonicus by extration with hot steam water (OJPl). The OIPl fraction was further purified by Sephadex G-50 gel filtration chromatography to produce FI (polysaccharides) and FII (oligosaccharides) fraction. The average molecular masses o fFI and FII fraction were determined to be 3050 kDa and 13 kDa, respectively. The antimicrobial activity of OIPl was tested against 8 strains of bacteria and one strain of yeast by the disc diffusion method, fluorescein diacetate (FDA) method and broth dilution method. The OIPl exhibited a very strong growth inhibition to Candida albicans. The OIPl remarkably sup­pressed the growth of Salmonella typhimurium and Staphylococcus aureus. The OIPl showed higher growth inhibition to Escherichia coli and Pseudomonas aeruginosa than propolis, positive control. When the anticancer activity of the OIPl, FI or FII was examined against human cancer cell lines and the Sarcoma 180 cells, these widely suppressed the proliferation of cell lines in the MTT assay and morphology study. Especially, they remarkably inhibited the growth of A549, HeLa and AGS cells. Also treatment of cancer cells with OJPl, FI or FII induced apoptotic cell death characterized by DNA fragmentation. The OJPl, FI or FII exhibiting various biological activities such as antimicrobial activity and anticancer activity is expected to be developed as new biohealth products.

Comparison of Harboring the Resistance Gene and Disc Diffusion Susceptibility Test Result in Staphylococcus pseudintermedius from the Bacterial Dermatitis (세균성 피부염 개에서 분리된 Staphylococcus pseudintermedius에서 항생제 감수성 검사와 내성 유전자 획득의 비교)

  • Jang, Hye-Jin;Son, Hyoung-Won;Kang, Hyo-Min;Han, Jae-Ik;Na, Ki-Jeong
    • Journal of Veterinary Clinics
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    • v.32 no.2
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    • pp.158-161
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    • 2015
  • Bacterial dermatitis is common disease that is necessary to treat with antibiotics. In recent, antibiotic-resistant bacteria is being increased in worldwide. The purpose of the present study was to evaluate the prevalence of resistant genes in Staphylococcus (S.) pseudintermedius isolated from dogs, and to compare the resistant gene profile with the result of antibiotic disc diffusion test. A total of seven S. pseudintermedius was included in the study. Bacterial identification was performed by 16S ribosomal RNA gene sequence analysis. S. pseudintermedius isolates had more than one antibiotic resistant gene (mecA, blaZ and aac(6')/aph(2"). While all isolates were PCR positive to blaZ gene, only two isolates were resistant to amoxicillin/clavulanate. Among five isolates harboring gentamicin resistance, one isolate was negative to aac(6')/aph(2")-targeted PCR. Taken together, the results suggest that resistant gene-targeted PCR and disc diffusion test are complementary to detect antibiotic resistance.

Distribution and Characterization of Integrons in Enterobacteriaceae Isolates from Chickens in Korea

  • Sung, Ji Youn;Oh, Ji-Eun
    • Journal of Microbiology and Biotechnology
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    • v.24 no.7
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    • pp.1008-1013
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    • 2014
  • The use of antimicrobial agents for additives or therapeutics is strongly associated with a prevalence of antimicrobial resistance in commensal Enterobacteriaceae. We aimed to characterize integrons in Enterobacteriaceae isolates obtained from chicken cecums in Korea. Moreover, the correlation between integron gene cassettes and antimicrobial resistance was also investigated. A total of 90 isolates the belonged to Enterobacteriaceae were recovered from chickens grown at Gyeongsang and Chungcheong provinces in Korea. Antimicrobial susceptibility tests were performed by the disk diffusion method. PCR and DNA sequencing were also performed to characterize the gene cassette arrays of the integrons. Of the 90 Enterobacteriaceae isolates tested, 39 (43.3%) and 10 (11.1%) isolates carried class 1 and 2 integrons, respectively. Whereas the class 2 integron did not contain gene cassettes, the class 1 integrons carried seven different gene cassette arrays. The class 1 integrons harbored genes encoding resistant determinants to aminoglycosides (aadA1, aadA2, and aadA5), trimethoprim (dfrA1, dfrA12, dfrA17, and dfrA32), lincosamides (linF), and erythromycin (ereA). Moreover, the presence of a class 1 integron was significantly related to a high resistance rate of antimicrobial agents, such as spectinomycin and trimethoprim. We confirmed that diverse class 1 integrons were widely distributed in Enterobacteriaceae isolates from chickens and directly contributed to the resistance to diverse antimicrobial agents in Korea.

Antagonism against Helicobacter Pylori and Proteolysis of Lactobacillus Helveticus CU631 and Strain Identification

  • Yoon, Y.H.;Won, B.R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.7
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    • pp.1057-1065
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    • 2002
  • The antagonistic activities of 30 strains of lactobacilli against Helicobacter pylori were determined and Lactobacillus helveticus CU631 has been selected as the strain which possesses the strongest inhibitory effect in the disc diffusion assay showing inhibition zone diameter of $10{\pm}1.5mm$, whereas those of L. plantarum and L. fermentum have been shown to be $4.0{\pm}0.6mm$. H. pylori G88016 revealed the highest vacuolating toxin producing activity among the 8 strains, the inhibitory activity of L. helveticus CU631 in vacuolating toxin producing activity of H. pylori manifested in the co-culture of two strains and in the 5:5 mixture of supernatant of the two strains. Both L. helveticus CU631 and cell free culture supernatant had a strong inhibitory activities in urease and cytotoxin producing activities of H. pylori NCTC11637 and CJH12. An accelerated proteolytic activity of water soluble peptides by L. helveticus CU631 during the refrigeration storage has been manifested in the cream cheese. DNA seqences of 16S-23S ribosomal RNA spacer region showed typical pattern among the various strains of L. helveticus, which could be used in the identification of L. helveticus CU 631.

Biochemical characterization and PFGE pattern of Brucella canis isolated from kennels in Gyoengbuk province (경북지역 애견 번식장에서 분리한 Brucella canis의 생화학적특성 및 PFGE 양상)

  • Kim, Seong-Guk;Kim, Young-Hoan;Hong, Hyon-Pyo;Eom, Hyun-Jung;Jang, Seong-Jun;Jo, Min-Hee;Lee, Yang-Soo
    • Korean Journal of Veterinary Service
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    • v.30 no.3
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    • pp.363-374
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    • 2007
  • A biochemical characterization and antimicrobial drugs susceptibility study was conducted in four breeding kennel which was canine abortion caused by Brucella canis in Gyeongbuk province in 2003-2006. Total of 267 dogs domesticated in the four kennel were examination. Among them, 143 (53.6%) dogs were sero-positive and 25 of blood samples were isolated to Brucella canis. At amplification of 35KDa-BCSP gene using PCR, 711 bp DNA fragment was same visible in 25 isolates and B canis RM6/66. Biochemical characterization of B canis isolated was non-hemolytic, no production of $H_2S$, no fermentation of carbohydrates, catalase-positive, oxidase-positive, indol-negative, hydrolyzation of urea, reduction of nitrate and development of thionin dye medium. Using disk-diffusion method, all of 25 strains tested were found to be highly susceptible to tetracycline, aminoglycoside, quinolone, macrolide antibiotics, rifampin and ampicillin in vitro. Using PFGE with restriction enzyme Smi I, 25 isolates tested were typed to 2 pattern, S1 and S2.