• Title/Summary/Keyword: Screened soil

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Ketoprofen ethyl ester에 대해 높은 광학 선택성을 갖는 (R)- 과 (S)-stereospecfic esterase들의 클로닝과 서열분석 및 발현

  • Kim, Ji-Yeon;Choe, Gi-Seop;Kim, Geun-Jung;Yu, Yeon-U
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.625-628
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    • 2001
  • To isolate novel strains that hydrolyzed the rac-ketoprofen ethyl ester to ketoprofen in the stereospecific manner, we screened broad ecological niches and soil samples in which the activity was expected to be found. From thousands of strains, we isolated a Pseudomonas sp. S34 producing a (S)-stereospecific esterase, and a thermostable esterase with (R)-form selectivity was also 。 btained from Bacillus stearothermophilus JYl44. To further analyse the gene structure and to induce a high level expression, two genes from each strain were cloned and sequenced. BLAST search results with the esterase gene from 534 revealed that both of gene (80-84 %) and amino acid sequences (89- 95 %) were highly conserved in the related esterases from Pseudomonas strains (fluorescens and aeruginosa). The thermostable esterase from JY144, however, revealed a relative low homology (45-52 %) to other esterase and/or lipase from related strains. Obviously, a complete conversion with pure enantiomer (R - or S) were readily achieved by recombinant clones expressing either (R)- or (S)- stereospecific esterase.

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Pathogenic and Molecular Characteristics of Agrobacterium vitis strains isolated from Grapevine in Korea

  • Kim, J.G.;Kim, S.H.;Choi, J.E.;Lee, Y.K.;Kang, H.W.
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.120.2-120
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    • 2003
  • Agrobacterium vitis is a causal agent of crown-gall disease on grapevine. In Korea, grapevine variety (GeoBong) have severely been infected by the bacteria since stems of the variety were buried in soil for overwintering. Infection ratio over 70-80% was observed on 7 years old GeoBong grapevine in Ansung and Cheonan. PCR specific primers for A. vitis strains were designed using nucleotide sequences of vir A gene in Ti-Plasmid, pheA gene in chromosomal DNA and a URP-PCR polymorphic band. Three hundred bacterial strains were isolated from the different 80 galls formed on GeoBong grapevine in Cheonan and Ansung of Korea and were screened to identify A. vitis using the three specific PCR primers for Agrobacterium vitis. Twenty-four bacterial strains that are detected by the primers were further confirmed by pathogenicity and biochemical methods. To investigate the genomic diversity of the bacterial strains, twenty primers of 20 mer referred to universal rice primers (URP) were applied for PCR fingerprinting, Of them, URP2R and URP2F primers could effectively be used to detect polymorphism within the bacterial strains.

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Cloning of \beta-mananase gene from Aeromonas sp. in E. coli (토양에서 분리한 Aeromonas sp 로 부터 \beta-mannanase 유전자의 클로닝)

  • 박봉환;강대경;김하근
    • Microbiology and Biotechnology Letters
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    • v.29 no.4
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    • pp.201-205
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    • 2001
  • A bacteria strain producing extracellular $\beta$-mannanase was isolated from soil and was identified as Aeromonas sp. A genomic DNA library constructed from Aeromonas, sp that secrets a $\beta$-mannanase was screened for mannan hydrolytic acticity. Recombinant $\beta$-mannanase activity was detercted on the basis of the clear zones around Escherichia coli colonies grown on a LB medium supplemented locust bean gum, EcoRI restriction analysis of plasmid prepared from recombinant E. coli which showed a $\beta$-mannanase activity revealed 10 kb DNA insert, The optimum pH and temperature for the activity of reconmbinant $\beta$-mannanase were 6.0 and $50^{\circ}C$ respectively and were identical to those of the native enzyme.

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Tn5 lac Mediated Mutagenesis of Enterobacter sp. B54 Antagonistic to Phytophthora capsici. (Phytophthora capsici의 성장을 저해하는 Enterobacter sp. B54의 선발과 Tn5 lac을 이용한 돌연변이 유기)

  • Yoon, Sang-Hong;Choi, Chung
    • Microbiology and Biotechnology Letters
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    • v.26 no.5
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    • pp.393-399
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    • 1998
  • Enterobacter sp.B54 which shows antagonistic activity to Phytophthora capsici on potato dextrose agar was selected among 112 strains isolated from Korean soil. After Tn5 lac-induced mutants were obtained through Pl :: Tn5 lac mutagenesis, 2 mutants for loss of antibiosis and 1 mutant for increased antibiosis were screened by using in vitro fungal inhibition assay. When the 3 mutants affected in antibiosis were analyzed by southern hybridization with pRZ102 (ColEl :: Tn5) as a probe, its results suggest that Tn5 lac was randomly inserted into different chromosomal sites in these mutants.

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A Newly Isolated Rhizopus microsporus var. chinensis Capable of Secreting Amyloytic Enzymes with Raw-Starch-Digesting Activity

  • Li, Yu-Na;Shi, Gui-Yang;Wang, Wu;Wang, Zheng-Xiang
    • Journal of Microbiology and Biotechnology
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    • v.20 no.2
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    • pp.383-390
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    • 2010
  • A newly isolated active producer of raw-starch-digesting amyloytic enzymes, Rhizopus microsporus var. chinensis CICIM-CU F0088, was screened and identified by morphological characteristics and molecular phylogenetic analyses. This fungus was isolated from the soil of Chinese glue pudding mill, and produced high levels of amylolytic activity under solid-state fermentation with supplementation of starch and wheat bran. Results of thin-layer chromatography showed there are two kinds of amyloytic enzymes formed by this strain, including one $\alpha$-amylase and two glucoamylases. It was found in the electron microscope experiments that the two glucoamylases can digest raw corn starch and have an optimal temperature of $70^{\circ}C$. These results signified that amyloytic enzymes secreted by strain Rhizopus microsporus var. chinensis CICIM-CU F0088 were types of thermostable amyloytic enzymes and able to digest raw corn starch.

Topical Anti-inflammatory Activity of Dianemycin Isolated from Streptomyces sp. MT 2705-4

  • Lee, Song-Jin;Kim, Hyun-Pyo;Park, Byung-Keun;Ahn, Soon-Cheol;Lee, Hyun-Sun;Ahn, Jong-Seog
    • Archives of Pharmacal Research
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    • v.20 no.4
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    • pp.372-374
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    • 1997
  • In order to develop new anti-inflammatory agents having different action mechanisms compared with nonsteroidal and steroidal anti-inflammatory drugs, the culture broths of various actinomycetes isolated from soil were screened using an in vivo mouse ear edma assay and one strain (Streptomyces sp. MT 2705-4: KCTC 8651 P) was selected. Activity-guided purification led to the isolation of a polyether compound, dianemycin. Topically, dianemycin showed a potent anti-inflammatory activity in mouse ear edema induced by croton-oil or arachidonic acid.$ED_{50}$value of dianemycin was found to be 0.8 mg,/ear compared to 0.4 mg/ear of prednisolone in croton-oil ear edema. However, dianemycin did not show the inhibitory activity in UV-erythema and delayed hypersensitivity reaction. These results indicate that dianemycin is a potential topical anti-inflammatory agent.

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Chitosanase를 생산 ${\cdot}$ 분비 하는 Bacillus sp. HSB-21의 분리 및 효소 특성

  • Kim, Seong-Gyun;Song, Hui-Sang;Kim, Dong-Seong;Sin, Jung-Han;Bang, Won-Gi;Choe, Yong-Bok
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.563-566
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    • 2000
  • In order to obtain microbial endochitosanase for enzymatic production of chitooligosaccharides from chitosan, we screened four microbes from soil and selected. Bacillus sp. HSB-21 which showed highest activity. Chitosanase, produced from isolating microbe, was endo-type and molecular mass of the enzyme was estimated as 21,000 by active staining. Its optimum pH and temperature were 5.5 and $50^{\circ}C$, respectively. It was stable in the pH range of 3.0 to 8.0 and up to $40^{\circ}C$. It did not produce chitomonosaccharide and produced chitooligosaccharide ranging from chitobiose to chitooctaose as major end-products from chitosan. The chitosanase from Bacillus sp. HSB-21 can be applicable to enzymatic production of chitooligosaccharide which has high degree of polymerization .

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Growth Promoting Rhizospheric and Endophytic Bacteria from Curcuma longa L. as Biocontrol Agents against Rhizome Rot and Leaf Blight Diseases

  • Vinayarani, G.;Prakash, H.S.
    • The Plant Pathology Journal
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    • v.34 no.3
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    • pp.218-235
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    • 2018
  • Plant growth promoting rhizobacteria and endophytic bacteria were isolated from different varieties of turmeric (Curcuma longa L.) from South India. Totally 50 strains representing, 30 PGPR and 20 endophytic bacteria were identified based on biochemical assays and 16S rDNA sequence analysis. The isolates were screened for antagonistic activity against Pythium aphanidermatum (Edson) Fitzp., and Rhizoctonia solani Kuhn., causing rhizome rot and leaf blight diseases in turmeric, by dual culture and liquid culture assays. Results revealed that only five isolates of PGPR and four endophytic bacteria showed more than 70% suppression of test pathogens in both assays. The SEM studies of interaction zone showed significant ultrastructural changes of the hyphae like shriveling, breakage and desication of the pathogens by PGPR B. cereus (RBacDOB-S24) and endophyte P. aeruginosa (BacDOB-E19). Selected isolates showed multiple Plant growth promoting traits. The rhizome bacterization followed by soil application of B. cereus (RBacDOB-S24) showed lowest Percent Disease Incidence (PDI) of rhizome rot and leaf blight, 16.4% and 15.5% respectively. Similarly, P. aeruginosa (BacDOB-E19) recorded PDI of rhizome rot (17.5%) and leaf blight (17.7%). The treatment of these promising isolates exhibited significant increase in plant height and fresh rhizome yield/plant in comparison with untreated control under greenhouse condition. Thereby, these isolates can be exploited as a potential biocontrol agent for suppressing rhizome rot and leaf blight diseases in turmeric.

Characterization and Culture Optimization of Regiospecific Cyclosporin Hydroxylation in Rare Actinomycetes Species

  • PARK, NAM-SIL;MYEONG, JI-SEON;PARK, HYUN-JOO;HAN, KYU-BOEM;KIM, SANG-NYUN;KIM, EUNG-SOO
    • Journal of Microbiology and Biotechnology
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    • v.15 no.1
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    • pp.188-191
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    • 2005
  • Abstract Cyclosporins are a family of clinically-important immunosuppressive cyclic peptides produced by Tolypocladium inflatum. The structural modification of cyclosporins via hydroxylation at various positions of N-methyl leucines in cyclosporin A leads to a dramatic change of their bioactive spectra. Among over 100 soil actinomycetes screened, two actinomycetes species, Sebekia benihana and Pseudonocardia autotrophica, were identified to contain superior cyclosporin A hydroxylation activities. A HPLC-based cyclosporin A hydroxylation assay revealed that each strain possesses distinctive hydroxylation specificity and regiospecificity; mono-hydroxylation at the 4th N-methyl leucine of cyclosporin A by S. benihana, and di-hydroxylations at both 4th and 9th N-methyl leucines of cyclosporin A by P. autotrophica. The conversion yields for cyclosporin A hydroxylation by both S. benihana and P. autotrophica were significantly improved from less than 10% and 18% up to 58% and 45%, respectively, in the optimized culture containing molybdenum with 0.05 g/l of cyclosporin A concentration. An ancymidol-specific inhibition of cyclosporin hydroxylation also suggested that the regiospecific cyclosporin hydroxylation might be catalyzed by a putative cytochrome P450 mono-oxygenase enzyme.

A Herbicidal Nucleoside Compound isolated from Streptomyces tubercidicus ME-9189 (Streptomyces tubercidicus ME-9189 균주가 생산하는 nucleoside계 제초 활성 물질)

  • Kim, Won-Gon;Kim, Jong-Pyung;Kim, Chang-Jin;Yoo, Ick-Dong
    • Microbiology and Biotechnology Letters
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    • v.24 no.1
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    • pp.82-86
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    • 1996
  • Three thousand microbial strains collected from different sources were screened for herbicidal activity. A strain of ME-9189 showed herbicidal activity against Digitaria sanguinalis and Portulaca oleracea was isolated from a mountainy soil. Based on taxonomic studies, the strain was identified as Streptomyces tubercidicus. The active compound of ME-9189 was purified from the culture broth by charcoal, silica gel, sephadex LH-20 column chromatography and crystalization, consecutively. The ME-9189 compound was identified as tubercidin by spectroscopic methods of UV, $^{1}H$ and $^{13}C$-NMR, and EIMS. In the bioassay, growth of radish shoot and root was inhibited by 50% with tubercidin treatment of 10 ppm, showing 2 times higher activity than that of herbicidin A and similar to that of toyocamycin.

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