• Title/Summary/Keyword: soil strain

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Production of D-Lactic Acid from DL-Lactonitrile by Pseudomonas sp. (Pseudomonas sp.에 의한 DL-Iactonitrile로부터 D-lactic acid의 생산)

  • 김현수;황인균;정남현;방원기
    • Microbiology and Biotechnology Letters
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    • v.30 no.4
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    • pp.373-379
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    • 2002
  • By using DL-acetonitrile as enzyme inducer, 90 bacteria were isolated from a field soil. Among the isolated strains, the strain WJ-003 showed the highest activity for production of D-lactic acid from DL-lactonitrile, and was partially identified as Pseudomonas sp. The production condition of D-lactic acid from DL-lactonitrile using resting cells as an enzyme source was optimized as follows: the reaction mixture contained 10 mM of DL-lactonitrile, 20 g of wet cells in 11 of 20 mM potassium phosphate buffer (pH 7.0) and the reaction was carried out at $30^{\circ}C$. After 18 h of reaction, 0.843 g/l of D-lactic acid was produced which corresponded to a conversion ratio of 93.7% and an optical purity of 99.8%. Additionally, when 10 mM of DL-lactonitrile was added once more to the reaction mixture at 14 h, 1.64 g/1 of D-lactic acid was produced after 28 h. In this experiment, the conversion ratio was 91.1% and optical purity 99.8%.

Cloning and Characterization of Pseudomonas mucidolens Exoinulinase

  • Kwon, Young-Man;Kim, Hwa-Young;Choi, Yong-Jin
    • Journal of Microbiology and Biotechnology
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    • v.10 no.2
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    • pp.238-243
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    • 2000
  • An exoinulinase (${\beta}-D-fructofuranosidase$) gene was cloned by chromosome walking along the upstream region of the endoinulinase gene of Pseudomonas mucidolens isolated from soil. the exoinulinase gene consisted of an ORF of 0,506 bp encoding a polypeptide of 501 amino acids with a deduced molecular weight of 55,000. The exoinulinase produced by the recombinant Escherichia coli $DH5{\alpha}$ strain was also purified to homogeneity as determined by SDS-PAGE and a zymogram. The molecular weight of the purified exoinulinase according to both SDS-PAGE and gel filtration matched the deduced molecular weight of the protein described above, thereby indicating that the native form of the exoinulinase was a monomer. The purified enzyme hydrolyzed activity value of 2.0. Furthermore, no inulo-oligomers were liberated from the inulin substrate in the enzymatic reaction mixtures incubated for 90 min at $55^{\circ}C$. Taken together, these results indicate that the purified ${\beta}-D-fructofuranosidase$ was an exoinulinase. The pH and temperature optima of the exoinulinase were pH 6.0 and $55^{\circ}C$, respectively. the enzymehad no apparent requirement for a cofactor, and its activity was completely inactivated by $Ag^{+},{\;}Hg^{2+},{\;}and{\;}Zn^{2+}$. Kinetic experiments gave $K_m,{\;}V_{max},{\;}and{\;}K_{cat}$ values for inulin of 11.5 mM, 18 nM/s, and $72{\;}s^{-1}$, respectively. the exoinulinase was fairly stable in broad pH conditions (pH 5-9), and at pH 6.0 it showed a residual activity of about 70% after 4 h incubation at $55^{\circ}C$.

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Antimicrobial Activity of N-Acetyl-Phenylalanine Produced from Streptomyces sp. G91353 (Streptomyces sp. G91353이 생산하는 N-Acetyl-Phenylalanine의 항균활성)

  • Kwon, Oh-Sung;Park, Hae-Ryong;Yun, Bong-Sik;Hwang, Ji-Hwan;Lee, Jae-Chan;Park, Dong-Jin;Kim, Chang-Jin
    • Microbiology and Biotechnology Letters
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    • v.34 no.4
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    • pp.306-310
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    • 2006
  • For screening of the compounds exhibiting antimicrobial activities against the D-alanyl-D-alanine of Gram positive bacteria, approximately 2,500 actinomycetes isolated from soil were examined far antimicrobial activity. In consequence, we recently isolated the Streptomyces sp. G91353 strain produced an active compound, A91353, that inhibits the growth of Gram positive bacteria. A91353 was identified as N-acetyl-phenylalanine by various spectroscopic methods. The minimum inhibitory concentration (MIC) values of N-acetyl-phenylalanine on Gram positive bacteria such as Streptococcus pyogenes 308A, Streptococcus pyogenes 77A were determined as $50{\mu}g/ml$, respectively, but did not effect on Gram negative strains. These results indicate that N-acetyl-phenylalanine have an antimicrobial activity, which may be caused by the disturbance of D-alanyl-D-alanine synthesis.

Isolation of Streptomyces sp. KK565 as a Producer of ${\beta}-Amyloid$ Aggregation Inhibitor

  • Hwang, Sung-Eun;Im, Hyung-Min;Kim, Dong-Hoon;Shin, Hyun-Ju;Shin, Dong-Hoon;Park, Jeong-Eun;Jo, In-Ho;Kim, Chang-Jin;Yoo, Jong-Shin;Kang, Jong-Min;Lim, Dong-Yeon;Ahn-Jo, Snag-Mee;Kwon, Ho-Jeong
    • Journal of Microbiology and Biotechnology
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    • v.13 no.5
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    • pp.809-814
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    • 2003
  • ${\beta}-amyloid$ ($A{\beta}$) peptides from the proteolytic processing of ${\beta}-amyloid$ precursor protein (${\beta}-APP$) aggregates in the brain to form senile plaques, and their aggregation plays a key role in pathogenesis of Alzheimer's disease (AD). To isolate an active compound that has an $A{\beta}$ aggregation-inhibitory activity, 2,000 microbial metabolite libraries were screened based on their ability to inhibit $A{\beta}$ aggregation by using both Congo red and thioflavin T assays. As a result, a water-soluble fraction of a soil microorganism, KK565, showed a potent $A{\beta}$ aggregation-inhibitory activity. The strain was identified as Streptomyces species, based on the cultural and morphological characteristics, the presence of diaminopimelic acid in the cell wall, and the sugar patterns for the whole-cell extract. In addition, the purification of active principle resulted in identifying a heat-unstable protein responsible for the $A{\beta}$ aggregation-inhibitory activity.

Purification of Materials Produced by Amylocolatosis sp. and Anticancer Effect in Oral Cancer Model (Amylocolatosis sp.가 생산하는 항암물질의 정제 및 구강암 모델에 미치는 항암 효과)

  • Kim, Jung;Park, Young-Min;Lim, Do-Seon
    • Journal of dental hygiene science
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    • v.3 no.1
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    • pp.11-14
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    • 2003
  • A methylotrophic Actinomycetes strain, which produce the anti-oral cancer activity compound, was isolated from soil and estimated as Amylocolatosis sp. based on taxonomic studies. A methanol didn't have influence on the production of the anticancer compounds. These compound were isolated by ethylacetate extract, silica gel column chromatography, sephadex LH-20 column and reverse phase HPLC. The compounds were very stable under heat ($121^{\circ}C$), acid(pH 2.0) and alkali(pH 11.0) treatment. The cytotoxic effect of isolated anticancer compounds on various cancer cell lines such as A549, SNU-1, KB, L1210, and Sarcoma 180 was investigated by MTT assay method. And these produced compounds also showed the broad antimicrobial spectrum to test strains such as bacteria and yeast.

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A Study on Creep Behavior of Geosynthetics Considering Effect of Temperature and Confining Stress (온도 및 구속응력을 고려한 토목섬유의 크리프거동에 관한 연구)

  • 방윤경;김홍택
    • Journal of the Korean Geotechnical Society
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    • v.19 no.5
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    • pp.291-299
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    • 2003
  • The effect of temperature and soil confining stress on geosyntheic creep behaviour was studied by performing the temperature dependent confined creep tests for HDPE geogrid and geomembrane specimen. The visco-elastic creep coefficients of the geosynthetics were evaluated by the test results and it was proposed that the simple expressions for the instantaneous and limit creep strain of geosynthetics was considered as a function of temperature and confining stress on geosynthetics. Based on the time-temperature superposition principle, a master curve has been drawn for extrapolating tensile creep strains to longer time intervals(1$\times$10 $^7$min.∼1$\times$10$^{10}$min.). By using this master curves, the shift factors which can be used in establishing master curve considering confining stress on geosynthetics were carried out. Each tests was performed during 8,000∼12,000 min., with temperature ranging between 5$^{\circ}C$ and 4$0^{\circ}C$ and with confining stress ranging between 0 t/$m^2$ and 9 t/$m^2$.

The Evaluation of Damage Behaviour of Adjacent Structures in Urban Excavation (도심지 흙막이 굴착시 인접구조물의 손상 평가연구)

  • 김학문;황의석
    • Journal of the Korean Geotechnical Society
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    • v.19 no.6
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    • pp.351-361
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    • 2003
  • Brick wall models with window opening, length 1.844m $\times$ height 0.6m, representing 2-story building was constructed on a scale of 1/10 of actual size of brick building for the investigation of damage mechanism. Six settlement troughs presenting six stages of excavation were simulated by Peck(1969) and O'Rourke et al.(1976) methods. The results from the model tests using Peck(1969) and O'Rourke et al.(1976) method indicated that angular distortion of brick wall by O'Rourke et at. method was 21% greater than that of Peck method. Horizontal displacement by O'Rourke et al.(1976) was 24% greater than that of Peck. When the degree of building damage for the O'Rourke et al. method of settlement trough is plotted on the damage level graph(Boscardin & Cording, 1989), damage level becomes much more severe than the level obtained by peck's method. Also, building stiffness and soil-structure interface are considered important factors of expressing building damage.

Optimization of Culture Conditions for Extracellular Lipid Production from Rhodotorula glutinis K-501 (Rhodotorula glutinis K-501로부터 세포외지질 생산을 위한 배양 조건의 최적화)

  • 김의용;박평규;채희정
    • KSBB Journal
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    • v.13 no.1
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    • pp.58-64
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    • 1998
  • An extracellular lipid-producing strain, Rhodotorula glutinis K-501, was isolated from soil. The extracellular lipid produced by the cell was found to have good and stable emulsifying activity. Factors affecting the extracellular lipid production were studied to optimize culture conditions for maximum production. Sucrose and ammonium sulfate were selected as best carbon and nitrogen sources, respectively because they gave the highest concentration of product. The optimum C/N ratio was 50. Optimum concentrations of $KH_2PO_4,\; Na_2HPO_4, \;and\; CaCl_2 $were 3.5, 1.0, 0.75, and 0.1g/L, respectively. The optimum temperature and initial pH were determined as $22^{\circ}C$ and 7.0, respectively. When the batch culture was conducted with a sucrose concentration of 60g/L under the optimized conditions, extracellular lipid was produced to a concentration of 8.1g/L with a high production yield of 51.9% based on dry cell weight.

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Isolation and characterization of a cDNA encoding a mammalian cathepsin L-like cysteine proteinase from Acanthmoeba healui

  • Hong, Yeon-Chul;Hwang, Mi-Yul;Yun, Ho-Cheol;Yu, Hak-Sun;Kong, Hyun-Hee;Yong, Tai-Soon;Chung, Dong-Il
    • Parasites, Hosts and Diseases
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    • v.40 no.1
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    • pp.17-24
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    • 2002
  • We have cloned a cDNA encoding a cysteine proteinase of the Acanthamoeba healui OC-3A strain isolated from the brain of a granulomatous amoebic encephalitis patient. A DNA probe for an A. healui cDNA library screening was amplified by PCR using degenerate oligonucleotide primers designed on the basis of conserved amino acids franking the active sites of cysteine and asparagine residues that are conserved in the eukaryotic cysteine proteinases. Cysteine proteinase gene of A. healui (AhCPI) was composed of 330 amino acids with signal sequence, a proposed pro-domain and a predicted active site made up of the catalytic residues, $Cys^{25},{\;}His^{159},{\;}and{\;}Asn^{175}$. Deduced amino acid sequence analysis indicates that AhCPI belong to ERFNIN subfamily of C 1 peptidases. By Northern blot analysis. no direct correlation was observed between AhCPI mRNA expression and virulence of Acanthamoeba, but the gene was expressed at higher level in amoebae isolated from soil than amoeba from clinical samples. These findings raise the possibility that AhCPI protein may play a role in protein metabolism and digestion of phagocytosed bacteria or host tissue debris rather than in invasion of amoebae into host tissue.

Production of Herbicide-resistant Transgenic Plants from Embryogenic Suspension Cultures of Cucumber (오이의 배발생 현탁 배양세포로부터 제초제 저항성 형질전환 식물체 생산)

  • 우제욱;정원중;최관삼;박효근;백남긴;유장렬
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.1
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    • pp.53-58
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    • 2001
  • To develop herbicide-resistant cucumber plants (Cucumis sativus L. cv Green Angle) embryogenic suspension cultures were co-cultured with Agrobacterium tumefaciens strain LBA4404 carrying a disarmed binary vector pGA-bar. The T-DNA region of this binary vector contains the nopalin synthase/neomycin phosphotransferase Ⅱ (npt Ⅱ) chimeric gene for kanamycin resistance and the cauliflower 35S/phosphinothricin acetyltransferase (bar) chimeric gene for phosphinothricin (PPT) resistance, After co-cultivation for 48 h, embryogenic calli were placed on maturation media containing 20 mg/L PPT. Approximately 200 putatively transgenic plantlets were obtained in hormone free media containing 40 mg/L PPT. Northern blot hybridization analysis confirmed the expression of the bar gene that was integrated into the genome of five transgenic plants. Transgenic cucumber plants were grown to maturity. Mature plants in soil showed tolerance to the commercial herbicide (Basta) of PPT at the manufacturer's suggested level (3 mL/L).

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