• Title/Summary/Keyword: minimal broth

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Site-specific Disruption of Glyoxylate Bypass and Its Effect in Lysine-producing Corynebacterium lactofermentum Strain

  • Kim, Youn-Hee;Lee, Heung-Shick
    • Journal of Microbiology and Biotechnology
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    • v.6 no.5
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    • pp.315-320
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    • 1996
  • The role of glyoxylate bypass in a lysine-producing Corynebacterium lactofermentum strain was analyzed. Unlike the wild type, the strain expressed enzymes of glyoxylate bypass during growth in the fermentation broth containing glucose as the carbon source. To evaluate the importance of glyoxylate bypass in the strain, we disrupted chromosomal aceA by using a cloned fragment of the gene. Site-specific disruption of aceA which codes for the isocitrate lyase, the first enzyme of the bypass, was confirmed by Southern blot analysis. The aceA mutant strain completely lost isocitrate lyase activity and ability to grow in a minimal medium containing acetate as the sole carbon source. The mutant strain was similar to its parental strain in growth characteristics and produced comparable amounts of lysine in shake flasks containing glucose as the carbon source. The amount of oxaloacetate accumulated in the fermentation medium was similar for both strains, suggesting that expression of glyoxylate bypass does not necessarily lead to the increase in intracellular oxaloacetate. These data clearly demonstrate that glyoxylate bypass does not function as one of the routes of carbon supply for lysine production in the strain. It appears that the leakiness of the glyoxylate bypass in the strain might be the result of a secondary mutation which arose during previous strain development by random mutagenesis.

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Growth-Inhibitory Effects of Cuscuta japonica Choisy's and C. australis R.Be's Extracts against Propionibacterium acnes (새삼 (Cuscuta japonica Choisy) 및 실새삼 (C. australis R.Be) 추출물의 여드름 유발균 Propionibacterium acnes 증식 억제 효과)

  • Lee, Sung-Ha;Kim, Kyong-Shin;Suk, Kui-Duk
    • Korean Journal of Pharmacognosy
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    • v.35 no.4 s.139
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    • pp.375-379
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    • 2004
  • According to the traditional chinese herbal Cuscuta japonica Choisy and C. australis R.Be (original plant of Semen Cuscutae in Korea) have been used to cure pimple. In order to prove their bacteriostatic effects against Propionibacterium acnes, we performed the micro broth dilution method. The minimal inhibitory concentration (MIC) of C. japonica juice was 27.3 mg/ml, but C. australis juice showed no effect. The MIC of water extracted C. japonica seed and Wontosa and Bupjetosa were $960\;{\mu}g/ml$. The MIC of both water and ethanol extracts of C. australis seed was $9240\;{\mu}g/ml$. But ethanol extracts of C. japonicaseeds, Wontosa and Bupjetosa had no bacteriostatic activity. We also performed the disk diffusion test, C. japonica juice had prominent effect. Water extracts of C. japonica seed, Wontosa and Bupjetosa showed bacterio-static effect in 10 mg/ml, and C. australis seed in 10 mg/ml. The ethanol extract of C. japonica seed, Wontosa and Bupjetosa, and C. australis seed showed bacteriostatic effect in 20 mg/ml.

Antimicrobial Activity and Acute Toxicity of Natural Rutin (천연 Rutin의 항균효과와 급성독성에 미치는 영향)

  • Rym, Kyo-Hwan;Eo, Seong-Kug;Kim, Young-So;Lee, Chong-Kil;Han, Seong-Sun
    • Korean Journal of Pharmacognosy
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    • v.27 no.4
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    • pp.309-315
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    • 1996
  • As part of our search for less toxic antimicrobial agents from natural resources, antimicrobial activity of rutin isolated from Sophora japonica was tested in vitro against four kinds of gram positive bacteria and four kinds of gram negative bacteria by serial broth dilution method. Among eight kinds of bacteria tested, the antimicrobial activity of rutin was the most potent against Mycobacterium smegmatis showing MIC of $375\;{\mu}g/ml$. The acute toxicity of rutin was examined in mice and rats. and $LD_{50}$ value administered intraperitoneally in mice was 650 mg/kg(Confidence limit: 894-1,473 mg/kg). There were no significant changes in serum biochemical values and histopathological changes as compared with those of control after intraperitoneal administration with rutin in rats.

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Secretion of Bovine $\beta$-Casein by Saccharomyces cerevisiae

  • Chung, Kun-Sub;Rafael, F.R.;Oh, Sang-Suk;Richardson, T.
    • Journal of Microbiology and Biotechnology
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    • v.1 no.1
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    • pp.31-36
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    • 1991
  • Yeast expression plasmids containing an appropriate leader sequence and bovine $\beta$-casein cDNA were constructed to produce $\beta$-casein for the study of its functional characteristics. Two kinds of expression systems for $\beta$-casein were constructed using pCGYl444 as a precursor plasmid. This plasmid is a yeast-E. coli shuttle vector which contains the chelatin promoter. The plasmid pISB202 contains the invertase leader sequence and $\beta$-casein gene. The plasmid pDEB303 contains the original bovine $\beta$-casein leader sequence gene. These two plasmids were introduced into S. cerevisiae AB116 which is a strain deficient in the major yeast proteinases. Each clone was grown in minimal media for 24 h before induction by $CuSO_4$. The cells were thus grown under expression conditions. Both strains harbouring pISB202 and pDEB303 expressed bovine $\beta$-casein. The $\beta$-casein was detected using immunochemical staining after western blot. Secretion of $\beta$-casein was detected in the culture broth. The estimated amount of secreted $\beta$-casein was approximately 50 ${\MU}g$/l.

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Implications of SPION and NBT Nanoparticles upon In Vitro and In Situ Biodegradation of LDPE Film

  • Kapri, Anil;Zaidi, M.G.H.;Goel, Reeta
    • Journal of Microbiology and Biotechnology
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    • v.20 no.6
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    • pp.1032-1041
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    • 2010
  • The comparative influence of two nanoparticles [viz., superparamagnetic iron oxide nanoparticles (SPION) and nanobarium titanate (NBT)] upon the in vitro and in situ low-density polyethylene (LDPE) biodegradation efficiency of a potential polymer-degrading microbial consortium was studied. Supplementation of 0.01% concentration (w/v) of the nanoparticles in minimal broth significantly increased the bacterial growth, along with early onset of the exponential phase. Under in vitro conditions, ${\lambda}$-max shifts were quicker with nanoparticles and Fourier transform infrared spectroscopy (FTIR) illustrated significant changes in CH/$CH_2$ vibrations, along with introduction of hydroxyl residues in the polymer backbone. Moreover, simultaneous thermogravimetric-differential thermogravimetry-differential thermal analysis (TG-DTG-DTA) reported multiple-step decomposition of LDPE degraded in the presence of nanoparticles. These findings were supported by scanning electron micrographs (SEM), which revealed greater dissolution of the film surface in the presence of nanoparticles. Furthermore, progressive degradation of the film was greatly enhanced when it was incubated under soil conditions for 3 months with the nanoparticles. The study highlights the significance of bacteria-nanoparticle interactions, which can dramatically influence key metabolic processes like biodegradation. The authors also propose the exploration of nanoparticles to influence various other microbial processes for commercial viabilities.

Endophytic Diaporthe sp. ED2 Produces a Novel Anti-Candidal Ketone Derivative

  • Yenn, Tong Woei;Ring, Leong Chean;Nee, Tan Wen;Khairuddean, Melati;Zakaria, Latiffah;Ibrahim, Darah
    • Journal of Microbiology and Biotechnology
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    • v.27 no.6
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    • pp.1065-1070
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    • 2017
  • This study aimed to examine the anti-candidal efficacy of a novel ketone derivative isolated from Diaporthe sp. ED2, an endophytic fungus residing in medicinal herb Orthosiphon stamieus Benth. The ethyl acetate extract of the fungal culture was separated by open column and reverse phase high-performance liquid chromatography (HPLC). The eluent at retention time 5.64 min in the HPLC system was the only compound that exhibited anti-candidal activity on Kirby-Bauer assay. The structure of the compound was also elucidated by nuclear magnetic resonance and spectroscopy techniques. The purified anti-candidal compound was obtained as a colorless solid and characterized as 3-hydroxy-5-methoxyhex-5-ene-2,4-dione. On broth microdilution assay, the compound also exhibited fungicidal activity on a clinical strain of Candida albicans at a minimal inhibitory concentration of $3.1{\mu}g/ml$. The killing kinetic analysis also revealed that the compound was fungicidal against C. albicans in a concentration- and time-dependent manner. The compound was heat-stable up to $70^{\circ}C$, but its anti-candidal activity was affected at pH 2.

Susceptibilities of Methicillin-Resistant Staphylococcus aureus (MRSA) Isolated from Milk of Bovine Mastitis to Antibiotics Combined with Sulbactam (젖소 유방염에서 분리된 메티실린 내성 황색포도상구균에 대한 항생제와 Sulbactam 병합의 항균효과)

  • Yoo, Jong-Hyun;Han, Hong-Ryul;Park, Hee-Myung
    • Journal of Veterinary Clinics
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    • v.25 no.4
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    • pp.231-235
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    • 2008
  • Various kinds of antibiotic intramammary infusions are used for treatment of bovine mastitis. As antibiotic-resistant bacteria are increased, the therapeutic rate for bovine mastitis is decreased. The goal of this research is to detect significant synergic effects of combination of antibiotics with sulbactam, $\beta$-lactamase inhibitor, on methicilin-resistant Staphylococcus aureus (MRSA). We used 5 strains of MRSA isolated from bovine mastitis with clinical and subclinical signs. All of the bacteria isolated had resistance to oxacillin and showed multi-resistant patterns in the antimicrobial susceptibility tests. Minimal bactericidal concentrations of ampicillin, amoxicillin, cephalexin, ampicillin/sulbactam(2:1), amoxicillin/sulbactam (2:1), and cephalexin/sulbactam (1:1) were measured according to broth microdilution method suggested by National Committee for Clinical Laboratory Standards (NCCLS, M31-A2) to compare the synergic effects of sulbactam combination with each antibiotic alone. Ampicillin and amoxicillin showed synergic antibacterial activity to 4 and 3 respectively in 5 strains of MRSA in combination with sulbactam. This study demonstrates that ampicillin/sulbactam and amoxicillin/sulbactam can be therapeutic choices for mastitis associated with MRSA.

Antibacterial Activity and Synergism of Hydnocarpi Semen Extracts with Ampicillin or Oxacillin against Methicillin-resistant Staphylococcus aureus

  • Joung, Dae Ki;Shin, Dong Young;Kwon, Dong Yeul;Shin, Dong Won
    • Korean Journal of Plant Resources
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    • v.29 no.6
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    • pp.699-703
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    • 2016
  • Methicillin-resistant Staphylococcus aureus (MRSA) is a serious clinical and an urgent problem worldwide. Few new drugs are available against MRSA, because MRSA has the ability to acquire resistance to most antibiotics, which consequently increases the cost of medication. In the present study, the antibacterial activity of Hydnocarpi Semen was investigated. The most effective method is to develop antibiotics from the natural products without having any toxic or side effects. Therefore, there is a need to develop alternative antibacterial drugs for the treatment of infectious diseases. Five Clinical isolates (MRSA) were obtained from five different patients at Wonkwang University Hospital (Iksan, South Korea). The Other 2 strains were ATCC 33591 (Methicillin-resistant strain) and ATCC 25923 (Methicillin-susceptible strain). Antibacterial activity (Minimal Inhibitory Concentrations, MICs) was determined by broth dilution method, disk diffusion method, MTT test, and checkerboard dilution test. Antibacterial activity of n-hexane fraction was remarkable, and had a MICs ranging from $31.25-125{\mu}g/m{\ell}$. FICI values for HFH+AM and HFH+OX were 0.13-0.19 and 0.04-0.29, showing the increase of synergistic effect. When combined together, these antibacterial effects were dramatically increased.

Distribution of Vancomycin-resistant Enterococci Isolates Using a ChromID VRE Agar

  • Lee, Hyun;Yoon, In-Seon
    • Korean Journal of Clinical Laboratory Science
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    • v.45 no.1
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    • pp.1-4
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    • 2013
  • Vancomycin-resistant enterococci (VRE) have emerged as important healthcare-associated infection since last two decades. ChromID VRE agar (cIDVA) is useful for VRE rectal swab screening. We investigated all VRE were isolated on the cIDVA. A total of 363 rectal swabs of 85 patients to test VRE screening were inoculated into bile-esculin (B-E) broth with $6{\mu}g/mL$ vancomycin. After 24 hours incubation, we subcultured B-E broths were changed to black onto cIDVA. All isolates were identified by the MICROSCAN and VITEK2. The vanA gene and vancomycin minimal inhibition concentration (MIC) were detected by PCR and E-test respectively. 277 E. faecium (84.7%), 16 E. faecalis (4.9%), 25 E. avium (7.6%), 8 E. gallinarum (2.4%) and 1 E. raffinosus (0.3%) were isolated. 10.3% of VRE detected on cIDVA were other than E. faecium and E. faecalis that presented various color from colorless to pale violet. All isolates contained vanA and vancomycin MIC were > $256{\mu}g/mL$. VRE isolates other than E. faecium and E. faecalis should be objective to the contact precautions for healthcare-associated infection control if they possess vanA gene. Due to emerging enterococci carrying vanA such as E. avium, E. gallinarum, and E. raffinosus, VRE surveillance should be expanded to all isolates on chromogenic agar.

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Isolation and Drug Sensitivity of Gram-Negative Rods from Bovine Udder Infections (젖소 유방염(乳房炎)으로부터 Gram 음성간균(陰性桿菌)의 분리(分離) 및 약제감수성(藥劑感受性)에 관(關)한 연구(硏究))

  • Lee, Cha Soo;Park, Cheong Kyu
    • Korean Journal of Veterinary Research
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    • v.20 no.2
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    • pp.79-84
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    • 1980
  • One hundred and eighteen cultures of Gram-negative rods isolated from cases of clinical bovine mastitis during lactation were examined for distribution of specific types, and activity of several antimicrobial agents to the isolates was determined by two-fold tube dilution method employing sterile whole milk as fluid medium. Of the isolates, 59.2% were Escherichia coli. Most of the remaining isolates were Klebsiella pneumoniae and Enterobacter aerogenes. Most Gram-negative rods(89.8%) were isolated from acute local and chronic mastitis. The cases of peracute systemic form with a marked symptoms of toxemia were associated with Escherichia coli and Klebsiella pneumoniae. The minimal lethal concentrations (MLC) of gentamicin and oxolinic acid in sterile whole milk were 16-128 times higher than the MLC obtained in trypticase soy broth (TSB), while the MLC of ampicillin and tetracycline in milk increased 2-4 times compared with TSB. Of the drugs tested, gentamicin was the most active antibiotics with MLC of $100{\mu}g/ml$ in sterile whole milk against all of Gram-negative rods isolated from bovine udder infections.

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