• Title/Summary/Keyword: Soil Culture

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Purification and Characterization of Transglutaminase from a Newly Isolated Streptomyces platensis YK-2 (토양 방선균 Streptomyces platensis YK-2가 생산하는 Transglutaminase의 정제 및 효소학적 특성)

  • Ko, Hee-Sun;Kim, Hyun-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.6
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    • pp.801-806
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    • 2009
  • A species producing transglutaminase (EC 2.3.2.13) was isolated from forest soil and identified as Streptomyces platensis YK-2. The transglutaminase was purified from culture broth by 50% methanol precipitation, followed by successive chromatography on DEAE-Sephadex. The yield and purification-fold was 63.4% and 2.2-fold, respectively. The purified microbial transglutaminase (MTG) migrated as a single band of approximately 45 kDa upon sodium dodecyl sulfate polyacrylamide gel eletrophoresis. The isoelectric point determined by multichambered electrofocusing was pH $6.0{\sim}7.0$. The enzyme was strongly inhibited by $Hg^{++}$, but was activated by $Cd^{++}$, $Mg^{++}$, $Mn^{++}$, $Pb^{++}$ and reducing agents such as dithiothreitol and mercaptoethanol.

Trypsin Inhibitor from Streptomyces sp. ( Part 1) Isolation of microorganism and purification of the inhibitor (Streptomyces 속 균주가 생성하는 Trypsin Inhibitor (제1보) 균의 분리 및 저해물질의 정제)

  • Yi, Dong-Heui;Seu, Jung-Hwn
    • Microbiology and Biotechnology Letters
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    • v.10 no.4
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    • pp.275-281
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    • 1982
  • One strain of Streptomyces sp. (AS-707) isolated from soil was found to produce a biologically active substance that showed a strong inhibitory activity against proteolytic enzymes viz. trypsin, papain, $\alpha$-chymotrypsin, Azotobacter protease, and Bacillus pretense. The substance was separated from culture filtrate by ion exchange column chromatography using Amberlite IRC-50 and CM-cellulose column chromatography. It was found that the recovery yield was 26% as activity basis. The substance was stable in wide pH range from 2.0 to 12.0 at 37$^{\circ}C$, but it was unstable in alkaline pH values at 6$0^{\circ}C$. The activity was thermostable to give 90% activity compared to the intact sample when it was treated at pH5.6 at 10$0^{\circ}C$ for 2 hours.

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절수의 시기 및 방법의 차이가 수도의 생육수량과 기타 실용형질에 미치는 영향

  • 이창구
    • Journal of the Korean Professional Engineers Association
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    • v.1 no.1
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    • pp.18-25
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    • 1968
  • Higher yield in rice paddies is greatly dependent on adequately balanced and timely supply of water. A majority of rice paddy in Korea is generally irrigated by rainfall, but in many cases it has to be supplemented by artificial irrigation for optimum rice culture. Although the water requirement of rice plant is for higher Than that of athec crops, submerged condition of rice paddy is not Necessarily required. The moisture requirement of rice plant varies with its growing stages, and it is possible to increase the irrigation efficiency through reduction of water loss due to percolation in rice paddies. An experimental plots were set up by means of sandomized block design with three duplication; (a) All time submerged, (b) Economically controlled, and (c) Extremely controlled. Three different irrigation periods Were (a) Initial sage, (b) Inter-stage, and (c) Yast stage. The topsoil of the three plots were excavated to the depth of 30 cm and then compacted with clay of 6 cm thickness. There after, they were piled up with the excavated top soils, leveled and cored with clay of 6 cm thickness around footpath in order to prevent leakage. The results obtained from the experiments are as follows, 1. There is no difference among the three experiments plots in terms of physical and chemical conditions, soil properties, and other characteristics. 2. Culm length and ear length are not affected by different irrigation methods. 3. There is no difference in the mature rate and 1, 000 grain weight of rice for the three plots. 4. The control plot which was irrigated every three days shows an increased yield over the all the time submerged plot by 17.8 percent. 5. The clay lined plot whose water holding capacity was held 5 days long, needs only to be irrigated every 7 days. 6. The clay lined plot shows an increased yield over the untreated plot ; over all-the-time submerged plot by 18 percent ; extremely controlled plot by 18 percent, and economically controlled plot by 33 percent. 7. It may be saved in water requirement about one Thirds.

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Isolation and Characteristics of Polyhydroxyalkanoates Producing Pseudomonas sp. MBEL21 (신규 Pseudomonas sp. MBEL21 균주의 Polyhydroxyalkanoates 생산 특성)

  • 최종일;이승환;이상엽
    • Microbiology and Biotechnology Letters
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    • v.32 no.2
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    • pp.123-127
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    • 2004
  • Pseudomonas sp. MBEL21 was newly isolated from soil samples and found to accumulate medium-chain-length Polyhydroxyalkanoates(MCL-PHAs) using oleic acid as a sole carbon source. Among the various nutrient limiting conditions examined, including nitrogen, sulfur and phosphorus, only phosphorus limitation supported the accumulation of MCL-PHAs up to 15 wt% of dry cell weight in flask cultures. MCL-PHAs produced by Pseudomonas sp. MBEL21 was mainly composed of 3-hydroxy-5-cis-tetradecenoate. Fed-batch culture of Pseudomonas sp. MBEL21 by novel feeding strategies based on cell growth charcteristics was carried out under phosphorus limitation using oleic acid as a sole carbon source. The final cell concentration and PHA content of 82 g/L and 28 wt%, respectively, were obtained. Furthermore, PHA consisted of MCL-hydroxyalkanoates and 3-hydroxybutyrate could be produced using olive oil as a sole carbon source.

Characterization and Antifungal Activity from Soilborne Streptomyces sp. AM50 towards Major Plant Pathogens

  • Jang, Jong-Ok;Lee, Jung-Bok;Kim, Beam-Soo;Kang, Sun-Chul;Hwang, Cher-Won;Shin, Kee-Sun;Kwon, Gi-Seok
    • Korean Journal of Environmental Agriculture
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    • v.30 no.3
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    • pp.346-356
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    • 2011
  • BACKGROUND: Chemical fungicides not only may pollute the ecosystem but also can be environmentally hazardous, as the chemicals accumulate in soil. Biological control is a frequently-used environment-friendly alternative to chemical pesticides in phytopathogen management. However, the use of microbial products as fungicides has limitations. This study isolated and characterized a three-antifungal-enzyme (chitinase, cellulase, and ${\beta}$-1,3-glucanase)-producing bacterium, and examined the conditions required to optimize the production of the antifungal enzymes. METHOD AND RESULTS: The antifungal enzymes chitinase, cellulase, and ${\beta}$-1,3-glucanase were produced by bacteria isolated from an sawmill in Korea. Based on the 16S ribosomal DNA sequence analysis, the bacterial strain AM50 was identical to Streptomyces sp. And their antifungal activity was optimized when Streptomyces sp. AM50 was grown aerobically in a medium composed of 0.4% chitin, 0.4% starch, 0.2% ammonium sulfate, 0.11% $Na_2HPO_4$, 0.07% $KH_2PO_4$, 0.0001% $MgSO_4$, and 0.0001% $MnSO_4$ at $30^{\circ}C$. A culture broth of Streptomyces sp. AM50 showed antifungal activity towards the hyphae of plant pathogenic fungi, including hyphae swelling and lysis in P. capsici, factors that may contribute to its suppression of plant pathogenic fungi. CONCLUSION(S): This study demonstrated the multiantifungal enzyme production by Streptomyces sp. AM50 for the biological control of major plant pathogens. Further studies will investigate the synergistic effect, to the growth regulations by biogenic amines and antifungal enzyme gene promoter.

Isolation and Characterization of Bacillus sp. P16 Producing Extracellular Chitosanase (키토산분해효소를 생산 분비하는 Bacillus sp. P16의 선발 및 특성)

  • Jung, Mi-Ra;Jo, Yoo-Young;Chi, Yeon-Tae;Park, Ro-Dong
    • Applied Biological Chemistry
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    • v.40 no.5
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    • pp.369-374
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    • 1997
  • An endochitosanase-producing bacterium was isolated from soil and identified as a strain of Bacillus sp. The isolate was gram positive, rod shape $(0.4-0.6{\times}1.6-2.2{\mu}m)$, endospore-forming, catalase positive, and mobility positive, and grown at pH 4.5-11.0 and upto $42^{\circ}C$ in the medium containing 2% NaCl. RAPD analysis of the DNA purified from the strain was also performed, and the chitosanase-producing strain was named as Bacillus sp. P16. The culture supernatant of the strain showed strong liquefaction activity and rapidly decreased viscosity of chitosan solution. By TLC and HPLC, chitooligosaccharides of DP 2-7 were separated and identified from the enzyme hydrolyzates of chitosan. The chitosanase from Bacillus sp. P16 was thus regarded as an endo-splitting type.

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Purification of Three Extracellular Proteases from Alkalophilic Coryneform Bacteria TU-19 (호알칼리성 Coryeform bacteria TU-19가 생산하는 세종류의 균체외 단백질분해호소의 정제)

  • Choi, Myoung-Chul;Yang, Jae-Sub;Kang, Sun-Chul
    • Applied Biological Chemistry
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    • v.38 no.6
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    • pp.534-540
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    • 1995
  • Alkalophilic coryneform bacteria TU-19 isolated from soil extracellularly produced at least three proteases (Protease I, II, and III). Investigating the cultural conditions related to the enzyme production of this bacterial cell, the optimum pH and temperature were 10.0 and $30^{\circ}C$, respectively. In order to purify these enzymes from the 2 day culture broth ammonium sulfate fractionation, gel filtration and QAE-Sephadex column chromatography were performed step by step. And then these three proteases were purified to near homogeneity by judging from SDS-PAGE pattern, and had the molecular weights of 120, 80, and 45 kilodaltons, respectively. The optimum pH and temperature for the enzyme activity of Protease I and II were 10.5 and $45^{\circ}C$, respectively, and Protease II were 11.0 and $50^{\circ}C$. And the enzymes were completely inhibited by PMSF suggesting serine protease, but not affected by pCMB. 1,10-phenanthroline, IAA, and EDTA.

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Studies on Cellulase -Part 1. Isolation of Cellulase Forming Microorganisms and the Properties of Crude Enzymes- (Cellulase에 관(關)한 연구(硏究) -(제1보(第一報)) Cellulase 생성균(生成菌)의 분리(分離)와 조효소(粗酵素)의 제성질(諸性質)-)

  • Chung, Dong-Hyo
    • Applied Biological Chemistry
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    • v.11
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    • pp.109-117
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    • 1969
  • Out of some 400 strains of Microorganisms, cellulase forming organisms was isolated from night soil during the course of screening tests. Two strains, Ku-3371 and Ku-4383 were found capable of producing cellulase in the shaking culture. General properties of the crude enzyme were as the following results. 1. The optimum pH values on CMC-saccharifying, CMC-liquefying and filter paper disintegrating activities were 4.0 to 5.5. 2. The stable pH range was within 3.5 to 6.5, 3. The optimum temperature was $40-45^{\circ}C$, the thermal stability was below $50^{\circ}C$ except on paper disintegrating activity and completely inactivated at $70^{\circ}C$. 4. Dialyzed crude enzyme was activated by $Mn^{2+}\;and\;Co^{2+}$ repectively but $Hg^{2+}$ was strong inhibitor.

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Isolation, Identification and Enzyme Properties of a Bacterium producing Alkaline Protease (Alkaline protease를 생산하는 미생물의 분리, 동정 및 효소성질)

  • Shin, Kong-Sik;Kang, Sang-Mo;Ko, Jung-Youn
    • Applied Biological Chemistry
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    • v.43 no.3
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    • pp.169-173
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    • 2000
  • For the development of enzyme detergent capable of effectively washing at low temperature, a bacterium producing alkaline protease was isolated from soil samples, and properties of the enzyme were investigated. The selected strain was Gram negative, rod shape$(0.6{\sim}0.7{\times}1.3{\sim}2.6\;{\mu}m\;in\;size)$ and motile. It had the degradation activity of aesculin, gelatin and casein, and was catalase-positive. The cell wall components was meso-DAP, and G+C mole contents was 43.3%. From these results, the strain was identified as Acinetobacter sp. KN-27. The activity of alkaline protease by this strain peaked with 3,300 D.U/mL after 36 hours in the liquid culture at $40^{\circ}C$. The optimal pH and temperature of the enzyme were pH 9 and $60^{\circ}C$, respectively. Alkaline protease produced by Acinetobacter sp. KN-27 has shown two active bands on the electrophoresis of native gel.

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Occurrence of Crown Gall of Rose and Rose Cultivar-specific Resistance (장미 뿌리혹병 발생과 품종간 저항성 차이)

  • Han Kyung-Sook;Kim Won-Hee;Park Jong-Han;Lee Jung-Sup;Seo Sang-Tae
    • Research in Plant Disease
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    • v.12 no.2
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    • pp.75-80
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    • 2006
  • Crown gall on rose was observed in greenhouse during year 2003-2005. The disease incidence was up to 28.3% and the disease was the severer in hydrophonics culture than that in soil. The typical gall symptom occurred mainly on the root and crown resulting in poor foliage, stunting, and fewer blossoms. Sixty-three rose cultivars were inoculated with Agrobacterium. tumefaciens isolated from rose crown gall, to evaluate rose cultivar-specific resistance. The size of galls from inoculated rose stems was measured in a greenhouse test. Tumors formed in almost varieties of rose inoculated. Based on the frequency of tumor occurrence and weight of galls formed on the stem of rose, it was shown that 'Little Marble', 'Golden Gate' and 'Rosa Rox-ette' were extremely susceptible to crown gall. Some varieties such as 'Little Silver' appeared to be resistant to the crown gall.