• Title/Summary/Keyword: agar concentration

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Isolation of Cadmium-Tolerant Bacteria and Characterization of Cadmium Accumulation into the Bacteria Cell (카드뮴 내성균(耐性菌)의 분리(分離), 동정(同定)및 균체내(菌體內) 카드뮴 축적(蓄積) 특성(特性))

  • Cho, Ju-Sik;Han, Mun-Gyu;Heo, Jong-Soo
    • Korean Journal of Environmental Agriculture
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    • v.11 no.1
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    • pp.77-85
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    • 1992
  • Of the cadmium-tolerant 162 bacterial strains isolated from soils, river waters or active sludges of waste-water disposal plants in the Gyeongnam province a strain C1, which showed considerably higher growth rate in the agar plate containing 2000 ppm than any other strains isolated, was identified as a Pseudomonas putida or its similar strain when analyzed by taxonomical characteristics. Optimum pH and temperature for the growth of the P, putida were 7.0 and $30^{\circ}C$, respectively. This strain was resistant to antibiotics(ampicillin, chloramphenicol and streptomycin), and heavy metals(lithium, cupper, lead and zinc). This strain utilized salicylate, naphthalene or xylene as a sole carbon source. The rate of cadmium accumulation in P. putida cell was enhanced at low concentration of Cd in the growth media. The maximum cadmium absorption by this strain grown in 1 and l0ppm of Cd was respectively 78% and 60% 24 hrs after culture, but in 100 ppm Cd, 40% 48 hrs after culture. Addition of a non-ionic surfactant Triton X-100(0.1%) to the medium enhanced the accumulation of cadmium in the P. putida up to approximately 37%.

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Antifungal Activity of the Extracts from Veratrum album L. var. grandiflorum Max. (백려노근경(白藜蘆根莖) 추출물(抽出物)의 항진균작용(抗眞菌作用))

  • Lee, Jong-Hwa;Kim, Won-Ja;Song, Byung-Sook;Cho, Sun-Hee
    • The Korean Journal of Pharmacology
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    • v.9 no.2
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    • pp.1-6
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    • 1973
  • Although numerous drugs are available for the treatment of superficial fungi infections of skin, the clinical effects of the majority of such drugs are not satisfactory. In the hope of searching the effective drugs for superficial fungi infections, authors studied whether Veratrum rhizoma extracts had any effect on fungi, with water extract (VRWE), ethanol extract (VREE) and methanol extract (VRME) from Veratrum album L. var. grandiflorum Max. In in vitro studies, the spores of fungi were inoculated on Sabouraud's glucose agar media which contained three extracts of Veratrun rhizoma in each concentration of $500\;{\mu}g/ml$, $1,000\;{\mu}g/ml$ and $5,000\;{\mu}g/ml$ respectively, and the growth of the fungi were observed for 3 weeks. The species of the fungi used in these experiments were Epidermophyton floccosum, Microsporum canis, Microsporum nanum, Microsporum gypseum, Microsporum cookei, Trichophyton rubrum, Trichophyton mentagrophytes, Trichophyton tonsurans and Trichophyton verrucosum. The results of the studies were as follows: 1. The growth of M. canis, M. nanum, T mentagrophytes and T. tonsurans were slightly inhibited by VRWE $1,000\;{\mu}g/ml$, and with VRWE $5,000\;{\mu}g/ml$, the growth of E. floccosum, M. gypseum and T. rubrum were slightly inhibited, moderate inhibition on the growth of M. canis, M. nanum, M. cookei, T. mentagrophytes and T. tonsurans were showed by VRWE $5,000\;{\mu}g/ml$. 2. With $500\;{\mu}g/ml$ of VREE, the inhibition on growth of E. floccosum, M. nanum and M. gypseum were slight, however significant inhibition on the growth of M. canis, M. cookei, T. mentagrophytes, T. rubrum and T. tonsurans were observed. The growth of M. nanum and M. gypseum were moderately inhibited, and significant inhibition on the growth of E. floccosum, M. canis, M. cookei, T. mentagrophytes, T. rubrum and T. tonsurans were observed by VREE $1,000\;{\mu}g/ml$. By VREE $5,000\;{\mu}g/ml$, the growth of all tested fungi were significantly inhibited except T. verrucosuia being showed slight inhibition. 3. Significant inhibition on the growth of M. canis, T, mentagrophytes, T. rubrum and T. tonsurans were noted, and moderate inhibition of M. nanum, slight inhibition of E. floccosum and M. gypseum in growth were observed by VRME $500\;{\mu}g/ml$. The growth of E. floccosum, M. canis, M. nanum, M. cookei, T. mentsgrophytes, T. rubrum and T. tonsurans were significantly inhibited by VRME $1,000\;{\mu}g/ml$, and that of M. gypseum was moderate. With $5,000\;{\mu}g/ml$ of VRME, significant inhibition on the growth of E. floccosum, M. canis, M. nanum, M. gypseum, M. cookei, T mentagrophytes, T. rubrum and T. tonsurans were observed, and T. verrucosum was showed only slight inhibition. From the above results, it was found that the extracts of organic solvents from Veratrum rhizoma (VREE & VRME) exerted significant antifungal activity, and their effects were probably derived from the pharmacological action of steroidal alkaloids.

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Identification and Fermentation Characteristics of Lactic Acid Bacteria Isolated from the Fermentation Broth of Korean Traditional Liquor, Andong-Soju (안동소주 발효액으로부터 분리한 젖산 세균의 동정 및 발효 특성)

  • Bae, Kyung-Hwa;Shin, Kee-Sun;Ryu, Hee-Young;Kwon, Chong-Suk;Sohn, Ho-Yong
    • Microbiology and Biotechnology Letters
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    • v.35 no.4
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    • pp.310-315
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    • 2007
  • To investigate the effect of lactic acid bacteria in Andong-Soju fermentation and traditional nuruk maturation, several lactic acid bacteria were isolated from the Andong-Soju fermentation broth and traditionally matured nuruks using Lactobacilli MRS agar containing bromocresol purple. Among the isolated bacteria, ADS-L1 showed the highest lactic acid production and was dominant species in fermentation broth. Based on physiological characteristics and 16S rDNA sequencing results, the ADS-L1 was identified as Pediococcus acidilactici. The ADS-L1 grew well at $50^{\circ}C$, and under the acidic conditions at pH 4, whereas the ADS-L1 failed to grew by treatments of 12% (w/v) ethanol or 0.01N HCl. Considering the high temperature of nuruk above $50^{\circ}C$ during nuruk maturation and the high ethanol concentration of broth above 12% at the end-stage of Andong-Soju fermentation, these results suggested that the ADS-L1 is popular in matured nuruks and plays role in the early-stage of fermentation. Analysis of pH, brix, reducing sugar content, lactic acid production, and cell growth during the cultivation of ADS-L1 further suggested that the ADS-L1 may contribute the prevention of contamination by rapid and steady acidification of broth, and do not cause problems by rapid death at the end-stage of fermentation.

Improving the Chitinolytic Activity of Bacillus pumilus SG2 by Random Mutagenesis

  • Vahed, Majid;Motalebi, Ebrahim;Rigi, Garshasb;Noghabi, Kambiz Akbari;Soudi, Mohammad Reza;Sadeghi, Mehdi;Ahmadian, Gholamreza
    • Journal of Microbiology and Biotechnology
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    • v.23 no.11
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    • pp.1519-1528
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    • 2013
  • Bacillus pumilus SG2, a halotolerant strain, expresses two major chitinases designated ChiS and ChiL that were induced by chitin and secreted into the supernatant. The present work aimed to obtain a mutant with higher chitinolytic activity through mutagenesis of Bacillus pumilus SG2 using a combination of UV irradiation and nitrous acid treatment. Following mutagenesis and screening on chitin agar and subsequent formation of halos, the mutated strains were examined for degradation of chitin under different conditions. A mutant designated AV2-9 was selected owing to its higher chitinase activity. To search for possible mutations in the whole operon including ChiS and ChiL, the entire chitinase operon, including the intergenic region, promoter, and two areas corresponding to the ChiS and ChiL ORF, was suquenced. Nucleotide sequence analysis of the complete chitinase operon from the SG2 and AV2-9 strains showed the presence of a mutation in the catalytic domain (GH18) of chitinase (ChiL). The results demonstrated that a single base change had occurred in the ChiL sequence in AV2-9. The wild-type chitinase, ChiL, and the mutant (designated ChiLm) were cloned, expressed, and purified in E. coli. Both enzymes showed similar profiles of activity at different ranges of pH, NaCl concentration, and temperature, but the mutant enzyme showed approximately 30% higher catalytic activity under all the conditions tested. The results obtained in this study showed that the thermal stability of chitinase increased in the mutant strain. Bioinformatics analysis was performed to predict changes in the stability of proteins caused by mutation.

Investigation of Active Antifungal Compounds of Essential Oil from Chamaecyparis obtusa Against Dermatophytes, Microsporum canis and Trichophyton Mentagrophytes (피부사상균 Microsporum canis 및 Trichophyton mentagrophytes에 대한 편백정유의 항진균활성물질 탐색)

  • Park, Mi-Jin;Lee, Soo-Min;Gwak, Ki-Seob;Jeung, Eui-Bae;Chang, Je-Won;Choi, In-Gyu
    • Journal of the Korean Wood Science and Technology
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    • v.33 no.3 s.131
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    • pp.72-78
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    • 2005
  • The present study was conducted to evaluate the application of Chamaecyparis obtusa and to investigate potential utilization of essential oil from C. obtusa as plant-based medicine. The antifungal activity of essential oil from leaves and twigs of C. obtusa (Sieb. Et Zucc) was determined and the major components of active fractions against Microsporum canis (KCTC6591) and Trichophyton mentagrophytes (KCTC6077) were identified by GC/MS analysis. In treatment of essential oil from C. obtusa, the strain M. canis was more resistant than the other, T. mentagrophytes. In the agar diffusion assay, essential oil from C. obtusa inhibited hyphal growth of M. canis and T. mentagrophytes at the concentration of more than 5,000 ppm. The zones named B and C in the TLC assay of essential oil from C. obtusa showed antifungal activities. Among four sub-fractions of n-hexane extract from B and C zones, named as B-1, B-2, C-1 and C-2, the C-2 showed the highest antifungal activity. Instrumental GC/MS analysis for sub-fractions showed that a major component of C-1 was ${\alpha}$-terpineol as terpene alcohol, while C-2 contained sesquiterpenes such as elemol, cedrol and eudesmol.

Probiotic Potential of Plant-Derived Lactic Acid Bacteria with Antihypertensive Activity (항고혈압 활성을 가진 식물유래 젖산균의 생균제 특성)

  • Lee, Ye-Ram;Son, Young-Jun;Park, Soo-Yun;Jang, Eun-Young;Yoo, Ji-Yeon;Son, Hong-Joo
    • Journal of Environmental Science International
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    • v.25 no.6
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    • pp.789-798
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    • 2016
  • Lactic acid bacteria (LAB) are industrially important microorganisms for probiotics. The recent widespread application of LAB for preparation of functional food is attributable to the accumulating scientific evidence showing their beneficial effects on human health. In this study, we isolated and characterized plant-derived LAB that show angiotensin-converting enzyme (ACE) inhibitory and antioxidant activities. The selected strain K2 was isolated from Kimchi, and identified as Lactobacillus plantarum by 16S rRNA gene analysis. The strain grew under static and shaking culture systems. They were also able to grow in different culture conditions like $25^{\circ}C{\sim}37^{\circ}C$ temperature, 4~10 pH range and ~6% NaCl concentration. L. plantarum K2 was highly resistant to acid stress; survival rate of the strain at pH 2.5 and 3 were 80% and 91.6%, respectively. The strain K2 also showed high bile resistance to 0.3% bile bovine and 0.3% bile extract with more than 74% of survival rate. The cell grown on MRS agar plate containing bile extract formed opaque precipitate zones around the colonies, indicating they have bile salt hydrolase activity. The strain showed an inhibitory activity against pathogenic bacteria such as Escherichia coli, Staphylococcus aureus and Listeria monocytogenes; antibacterial activity was probably due to the lactic acid. The K2 strain showed relatively higher autoaggregation values, antihypertensive and antioxidant activities. These results suggest that L. plantarum K2 could be not only applied as a pharmabiotic for human health but also is also starter culture applicable to fermentative products.

Antimicrobial Characteristics of Different Parts of Guava against Food-Borne Bacteria (식중독 세균에 대한 구아바 부위별 추출물의 항균 특성)

  • Jo, Young-Hong;Ok, Dul-lee;Lee, Seung-Cheol
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.12
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    • pp.1773-1778
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    • 2009
  • Guava (Psidium guajava L.) contains high amount of vitamins and minerals, and its leaves have been reported to be very effective on reducing blood pressure. In this study, antimicrobial characteristics of extracts from four different parts of guava (fruit, branch, leaf, and seed) with four different solvents (methanol, ethanol, acetone, and water) were evaluated. Four targeted food-borne microorganisms were selected; two Gram negatives (Escherichia coli O157:H7 and Salmonella Typhimurium) and two Gram positives (Staphylococcus aureus and Listeria monocytogenes). By the paper disc method, guava extracts showed stronger clear hollow zone against Gram positive bacteria than Gram negatives in nutrient broth agar plate. Especially, extracts of branches and leaves showed significant antibacterial activity. Guava extracts also showed significant inhibition activity on the growth of Gram positive food-borne bacteria in nutrient broth. For example, S. aureus did not grow at all at 200 ppm of acetone extracts of guava branch and leaf. In the case of L. monocytogenes, the same concentration of acetone extracts of guava branch and leaf inhibited the growth 33.5% and 55.4%, respectively, at 32 hrs of incubation time. The results indicated that extracts of guava branch and leaf showed significant antibacterial activities against food-borne Gram positive microorganisms, and that guava branches, the byproducts of guava, might be a valuable resource for antibacterial materials.

Study on CsRCI2D and CsRCI2H for improvement of abiotic stress tolerance in Camelina sativa L.

  • Lim, Hyun-Gyu;Kim, Hyun-Sung;Kim, Jung-Eun;Ahn, Sung-Ju
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.196-196
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    • 2017
  • Oilseed crop Camelina (Camelina sativa L.) is a suitable for biodiesel production that has high adaptability under low-nutrient condition like marginal land and requires low-input cost for cultivation. Enhanced abiotic stress tolerance of Camelina is very important for oil production under the wide range of different climate. CsRCI2s (Rare Cold Inducible 2) are related proteins in various abiotic stresses that predicted to localized at plasma membrane (PM) and endoplasmic reticulum (ER). These proteins are consist of eight-family that can be divided into tail (CsRCI2D/E/F/G) and no-tail (CsRCI2A/B/E/H) type of C-terminal. However, it is still less understood the function of C-terminal tail. In this study, CsRCI2D/H genes were cloned through gateway cloning system that used pCB302-3 as destination vector. And we used agrobacterium-mediated transformation system for generation of overexpression (OX) transformants. Overexpression of target gene was confirmed using RT-PCR and segregation ratio on selection media. We analyzed physiological response in media and soil under abiotic stresses using CsRCI2D and CsRCI2H overexpression plant. To compare abiotic stresses tolerance, wild type and CsRCI2D/H OX line seeds were sown on agar plate treated with various NaCl and mannitol concentration for 7 days. In the test of growth rate under abiotic stress on media, CsRCI2H OX line showed similar to NaCl and mannitol stress. In the other hand, CsRCI2D OX line showed to be improved stress tolerance that especially increased in 200mM NaCl but was similar on mannitol media. In greenhouse, WT and CsRCI2D/H OX lines for physiological analysis and productivity under abiotic stresses were treated 100, 150, 200mM NaCl. Then it was measured various parameters such as leaf width and length, plant height, total seed weight, flower number, seed number. CsRCI2H OX line in greenhouse did not show any changes in physiological parameters but CsRCI2D OX line was improved both physiological response and productivity under NaCl stress. Among physiological parameters of CsRCI2D OX line under NaCl stress, leaf length and width were observed shorter than WT but it were slightly longer than WT in 200mM NaCl stress. Furthermore, total seed weight of CsRCI2D OX line under stress displayed to decrease than WT in normal condition, but it was gradually raised with increasing NaCl stress then more than WT relatively. These results suggested CsRCI2D might be contribute to improve abiotic stress tolerance. However, function of CsRCI2H is need to more detail study. In conclusion, overexpression of CsRCI2s family can generate various environmental stress tolerance plant and may improve crop productivity for bio-energy production.

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Lysine Production by Thialysine Resistant Mutant of Candida utilis ( I ) - Isolation of High Lysine Excreting Mutant of Candida utilis - (Candida utilis의 Thialysine 내성맥리주에 의한 Lysine생산 ( I ) -Candida utilis의 Lysine을 생산하는 Thialysine 내성맥리주의 분리-)

  • Bang, Byung-Ho;Seu, Jung-Hwn
    • Microbiology and Biotechnology Letters
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    • v.11 no.3
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    • pp.175-180
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    • 1983
  • Thialysine significantly inhibited the growth of wild type strain Gondida utilis NCYC-359. In the absence of thialysine, the culture reached stationary phase after 24hr, however, in the presence of 0.5% thialysine, the culture reached stationary phase after 40hr, respectively. Effect of amino acid or vitamin was investigated on recovery of the growth of wild type strain from thialysine inhibition. Glycine, methionine, arginine and tryptophan recovered growth inhibition by thialyslne to some extent. However, vitamins were inert. Especially, lysine at one eighth concentration of thialysine recovered almost fully the growth inhibition. Thialysine resistant mutants were induced from the parent strain of Condida utilis NCYC-359 by NTG treatment. Colonies of thialysine resistant mutants were obtained on agar minimal medium supplemented with 0.1-0.5% thialysine. The frequency of thialysine resistant mutants induced by the first mutation was the highest at 0.1% The wild strain produced no appreciable lysine extracellularly. However, almost thialysine resistant mutants excreted appreciably. Lysine excretion increased after repeated mutation. Finally, of the thialysine resistant mutants induced by NTG, Condida utilis TRN-4006 was obtained. This strain excreted lysine (400$\mu\textrm{g}$/$m\ell$) into the medium with a concomitant decrease of lysine in the intracellular pool.

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Detection and Distribution of the Pathogenic Bioagent Aeromonas (Gamma-Proteobacteria) in Water Supplies of Seoul (서울시 상수계통에서 병원성균 Aeromonas (감마-프로테오박테리아) 분포연구)

  • Lee, Eun-Sook;Lee, Mok-Young;Han, Sun-Hee;Ka, Jong-Ok
    • Korean Journal of Microbiology
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    • v.43 no.2
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    • pp.106-110
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    • 2007
  • The detection and distribution of Aeromonas in water supplies were investigated by using the USEPA Method 1605. Water samples were collected from the Han River, finished waters and tap waters supplied from Water Treatment Plants in Seoul monthly from July 2002 to December 2003. Aeromonas species in each water sample were quantified based on the development of yellow colonies on the surface of membrane filter using a selective medium (Ampicillin-Dextrin Agar with Vancomycin). The Quality Control (QC) for this study met the acceptance criteria of Method 1605. The concentrations of Aeromonas species in surface water samples ranged from $1.0{\times}10^{0}\;to\;9.8{\times}10^{3}\;CFU/ml$. Aeromonas species were found only in one tap water sample with concentration of 1 CFU/500 ml. No Aeromonas species were found in any finished water samples. Aeromonas species detected here were identified as A. salmonicida(51%), A. caviae(4.7%), A. schubertti(3.4%), A. sobria(3.8%), A. hydrophila(2.1%), and A. ichithiosmia(0.4%). A. salmonicida was the dominant species, which is of no significance to human health. Chlorine resistance of A. salmonicida was evaluated and as a result, 99.99% of A. salmonicida decreased after 30 seconds exposure at residual free chlorine 0.2 mg/L. These suggest that the waters supplied in Seoul may be safe against the pathogenic agent Aeromonas.