• Title/Summary/Keyword: 토양 효소 활성

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Purification and characterization of the chitinase from Bacillus subtilis JK-56 (Bacillus subtilis JK-56이 생산하는 chitinase isozyme의 정제와 특성 규명)

  • 전홍기;김낙원;정영기
    • Journal of Life Science
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    • v.12 no.1
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    • pp.77-86
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    • 2002
  • Chitin, a $\beta$-1,4 polymer of N-acetyl-D-glucosamine, is one of the most abundant organic compounds in nature. Chitinase (EC 3.2.1.14) is an enzyme that degrades chitin to chito-oligosaccharides, diacetyl rhitobiose and N-acetyl-D-glucosamine. An extracellular chitinase-producing bacterial strain was isolated from soil and named to as Bacillus subtilis JK-56. Optimum culture condition of B. subtilis JK-56 for the production of chitinase was 1% chitin, 0.5% polypepton, 0.1% KCl, 0.05% MnS $O_4$.4$H_2O$, 37$^{\circ}C$, initial pH 7.0 and 40 hour culture time. When B. subtilis JK-56 was grown in the optimum medium, one major active band and two minor active bands were detected by native-PAGE and active staining of the gel. Among them, the major band was purified from the culture supernatant by 70% ammonium sulfate precipitation and native-PAGE with BIO-RAD Model 491 Prep-Cell and named as Chi-56A. Its molecular weight was estimated to be 53kDa monomer and the isoelectric point (pI) was pH 4.3. The pH and temperature for the optimum activity of Chi-56A were pH 6.0 and $65^{\circ}C$, respectively. Chi-56A was stable up to $65^{\circ}C$ and in alkaline region. Its $K_{m}$ value for colloidal chitin was 17.33g/L. HPLC analysis of the reaction products confirmed that Chi-56A was an exo type chitinase.e.

Influence of Growing Green Manures on Soil Microbial Activity and Diversity under Organically Managed Grape-greenhouse (시설 포도 유기농 재배지에서 녹비 생육 중 녹비 종류가 토양 미생물상에 미치는 영향)

  • Park, Kee-Choon;Seo, Young-Jin;Kim, Chan-Yong;Kim, Jong-Su;Yi, Young-Keun;Seo, Ji-Ae
    • Korean Journal of Environmental Agriculture
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    • v.27 no.3
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    • pp.260-266
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    • 2008
  • The aim of present work was to assess the response of soil microbial activity and diversity to green manures under the organically managed grape-greenhouse in early spring. Hairy vetch, milk vetch, and red clover were seeded in fall, and enzymatic activities by dehydrogenase and fluorescein diacetate (FDA) hydrolase, and microbial diversities by Biolog $EcoPlate^{TM}$ and phospholipid fatty acid (PLFA) were characterized for soils sampled in early spring. Dehydrogenase activity and FDA hydrolytic activity did not differentiate the green manures but the average well color development of Biolog EcoPlate was higher in soils covered with red clover than control soil. Soil microbial functional diversity by Biolog EcoPlate differentiated the soils covered with hairy vetch and milk vetch, and Shannon diversity index by Biolog EcoPlate was higher in soils covered with hairy vetch than control soil. Principal component analysis of PLFA differentiated the soils covered with milk vetch from control soil.

Studies on the Microbial Glucose Isomerase Part 1. Isolation and Characterization of Streptomyces species Producing Glucose Isomerase (미생물의 포도당 이성화효소에 관한 연구 (제1보) 포도당 이성화효소 생산균주의 분리 및 성질에 관하여)

  • Chung, Tai-Wha;Kim, Hyun U.;Moon H. Han
    • Microbiology and Biotechnology Letters
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    • v.4 no.4
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    • pp.138-144
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    • 1976
  • five strains of Streptomyces spp. with high Productivity of glucose isomerase (15-30 units/ml) were obtained among 280 microbial strains isolated from 150 soil samples. These strains produced glucose isomerase with xylose as an inducer. These 5 strains were also identified to be different strains of Streptomyces spp.:streptomyces sp. K-14, K-53, K-71, K-77 and K-733. It was found that Streptomyces sp. K-14 produced the highest enzyme activity. The spore chains of these strains were rectiflexible and spore surface was smooth except Steptomyces sp. K-77 and K-733, with spiny surface.

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Influence of Long-term Fertilization on soil Enzymes Activity in Normal Paddy Soil (퇴비(堆肥) 및 비료(肥料) 장기연용(長期連用)이 토양내(土壤內) 효소활성(酵素活性)에 미치는 영향(影響))

  • Cho, Kang-Jin;Jung, Yeun-Tae;Choi, Jyung
    • Applied Biological Chemistry
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    • v.32 no.2
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    • pp.109-115
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    • 1989
  • This study was aimed to find out the influence of long-term fertilization for 21 years on soil enzyme activities in the silty clay loam textured normal paddy soil. Total urease activity (TUA) and the microbial urease activity (MUA) were shown to be changed significantly, but the accumulated urease activity (AUA) was similar within trial plots. Especially the MUA of the plots annually applied N.P.K. fertilizers with compost and N.P.K. fertilizers with silicate fertilizer were the highest among plots. The total L-glutaminase activity (TGA) and the accumulated L-glutaminase activity(AGA) were changed significantly among trial plots, but the microbial L-glutaminase activity (MGA) was not. By the simple correlation analysis, it was shown that the TGA and the AGA correlated highly significant to available phosphorus available $SiO_2$ content and pH. Addition of the toluene to the incubation mixture did not markedly affect the activity of phosphatase, but the difference of phosphatase activity among plots was significant. By the simple correlation analysis, it was shown that the phosphatase activity ; correlated highly significant to pH, available $SiO_2$, available phosphorus and exchangeable calcium in soils.

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Effect of activities of monooxygenase, ${\alpha}$, ${\beta}-esterase$ on the degradation of diazinon and dursban in submerged soil (담수토양중 Diazinon과 Dursban의 분해에 미치는 Monooxygenase와 Esterase의 활성)

  • Choi, Jong-Woo;Rhee, Young-Ha;Lee, Kyu-Seung
    • Korean Journal of Environmental Agriculture
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    • v.9 no.2
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    • pp.97-103
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    • 1990
  • In order to determine the major biochemical degradation factors of the two organophosphorus insecticides, diazinon and dursban, the activities of monooxygenase(m. o.) and ${\alpha}$, ${\beta}-esterase$ were studied in submerged soil under laboratory conditions at $30{\pm}1^{\circ}C$ The degradation rate of diazinon by microorganism showed 1.5 times higher than dursban. The m. o. activity increased from 12hrs and 3days after application of diazinon and dursban, respectively. But the ${\beta}-esterase$ activity showed maximum at one day after application of dursban and $5{\sim}8$ days after diazinon application. Also, the ${\beta}-esterase$ activity was about 10 times higher than ${\alpha}-esterase$. Hence, it was concluded that the biological degradation of diazinon was mainly attributed to m. o. activity and the degradation of dursban to ${\beta}-esterase$ activity.

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Purification and Characterization of Extracellular Inulinase from Bacillus sp. (Bacillus sp.가 세포외로 생산하는 Inulinase의 정제 및 특성)

  • 김경남;최용진
    • Microbiology and Biotechnology Letters
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    • v.18 no.5
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    • pp.490-495
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    • 1990
  • The extracellular inulinase from Bacillus spp. was purified to a single protein through a sequence of operations including ammonium sulfate fractionation, heat treatment, DEAE Sepharose C1-6B ion exchange chromatography, Sephadex 6-100 and Sephadex 6-150 gel filtration. The purified enzyme was confirmed to be a $\beta$ -D-fructofuranosidase(EC 3.2.1.26) which was much more active on sucrose than on inulin(I/S = 0.2). The maximal inulinase activity was observed at pH 6.0 and at the temperature of $50^{\circ}C$. The mo1ecular weight of the enzyme was about 56, 000. Tryptophan and histidine residues of the enzyme molecule were found to be essential for its catalytic activity.

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Effect of Soil surface Soil Management Practices on Microflora in Volcanic Ash Soils of Citrus Orchard (화산회토 감귤원의 표토관리방법이 토양 미생물상에 미치는 영향)

  • Joa, Jae-Ho;Lim, Han-Cheol;Koh, Sang-wook;Hyun, Hae-Nam
    • Korean Journal of Soil Science and Fertilizer
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    • v.37 no.3
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    • pp.165-170
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    • 2004
  • This study was conducted to investigate the effect of different surface soil management practices on soil microflora in volcanic ash soils of citrus orchard. Soil samples were collected from citrus orchards of clean cultivation, grass sod, and grass mulch system in May and September 1997. Soil chemical properties, populations of various microorganisms, enzyme activities, microbial biomass C were analyzed. Average soil pH were 4.7, and average nitrogen and organic matter contents were 6 and $140.2g\;kg^{-1}$, respectively. Aerobic bacteria were distributed at $26,2-47.3{\times}10^6cfu\;g^{-1}$ level. Among the aerobic bacteria Pseudomonas spp., Rhizobium spp., and thermophilic Bacillus spp. were dominant in most of the investigated orchard soils. Density of actinomycetes were low at $1.8-84.6{\times}10^5cfu\;g^{-1}$ level. Fungi were distributed at $26.4-182.1{\times}10^5cfu\;g^{-1}$ level and the density was higher in grass mulch and sward sites. In september, phosphomonoesterase activity was high at $239.6{\mu}g\;PNP\;g\;soil^{-1}\;h^{-1}$ in clean cultivated citrus orchards. Soil cellulase activity were higher at $602.6{\mu}g\;GE\;g\;soil^{-1}$\;24\;h^{-1}$ in grass sward cultivation than any other soil management practices. Soil microbial biomass C was higher in grass mulch cultivated orchards.

Partial purification and Properties of Alkaline Cellulase from Pseudomonas sp. AC-711 (Pseudomonas sp. AC-711이 생산하는 알칼리성 Cellulase의 부분정제 및 효소적 성질)

  • Yoon, Min-Ho;Lim, Sang-Ho;Choi, Woo-Young
    • Korean Journal of Agricultural Science
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    • v.25 no.1
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    • pp.131-137
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    • 1998
  • The cellulase components were partially purified from the culture filtrate of the alkalophilic bacterium Pseudomonas sp. AC-711 and its enzymatic properties were characterized. The specific activity of the purified major enzyme component was 3.5 units/mg protein as carboxymethyl cellulase and the yield was 23% of the total activity of the culture broth. The molecular weight of the component was 46,000 and the Km and Vmax on CMC were determined as $15.4mg\;mL^{-1}$ and $4.17{\mu}moles\;mL^{-1}\;min^{-1}$, respectively. The enzyme was stable at the temperatures below $60^{\circ}C$ and at the pH range of 4.0~11.0, and the optimal temperature and pH were $60^{\circ}C$ and pH 8.0, respectively. The enzyme activity was not significantly affected by the common surfactants (concentration: 0.05%) such as ${\alpha}$-olefin sulfonate, linear alkylbenzene sulfonate, sodium dodecyl sulfonate, hexadecyltrimethylammonium bromide and Tween 80. The enzyme was activated by the metal ions such as $Ca^{2+}$, $Cu^{2+}$, $Co^{2+}$, whereas inhibited by $Hg^{2+}$ and $Zn^{2+}$. The enzyme exhibited relatively high activity toward amorphous CMC as compared with crystalline substrates such as filter paper and avicel.

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Purification and Biochemical Characteristics of Fibrinolytic Enzyme from Streptomyces corcohrussi JK-20 (Streptomyces corcohrussi JK-20 유래 혈전용해효소의 순수분리 및 이의 생화학적 특성 규명)

  • Kim, You-Jung;Park, Jeong-Uck;Seo, Min-Jeong;Kim, Min-Jeong;Lee, Hye-Hyeon;Jin, Se-Hun;Kang, Byoung-Won;Choi, Yung-Hyun;Jeong, Yong-Kee
    • Journal of Life Science
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    • v.20 no.6
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    • pp.838-844
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    • 2010
  • A fibrinolytic enzyme of Streptomyces corcohrussi from soil sediment was purified by chromatography using DEAE-Sephadex A-50 and Sephadex G-50. The analysis of SDS-polyacrylamide gel suggested that the purified enzyme is a homogeneous protein and the molecular mass is approximately 34 kDa. The purified enzyme showed activity of 0.8 U/ml in a plasminogen-rich fibrin plate, while its activity in a plasminogen-free fibrin plate was only 0.36 U/ml. These results suggested that the purified enzyme acts as a plasminogen activator. The fibrinolytic activity of the enzyme under the supplementation of protease inhibitors, $\varepsilon$-ACA, t-AMCHA and mercuric chloride in the enzyme reaction was less than 24%, indicating that it could be modulated by the plasmin and/or fibrinogen inhibitors involved in the fibrinogen-to-fibrin converting process. As time passed, $Zn^{2+}$, a heavy metal ion, inhibited the activity to 34.1%. The optimum temperature of the purified enzyme was approximately $50^{\circ}C$ and over 92% of the enzyme activity was maintained between pH 5.0 and 8.0. Therefore, our results provide a potential fibrinolytic enzyme as a noble thrombolytic agent from S. corcohrussi.

Impacts of Chemical Properties on Microbial Population from Upland Soils in Gyeongnam Province (경남지역 밭 토양 화학성분이 미생물 생태에 미치는 영향)

  • Lee, Young-Han;Ha, Sang-Keun
    • Korean Journal of Soil Science and Fertilizer
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    • v.44 no.2
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    • pp.242-247
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    • 2011
  • Soil management for environment-friendly agriculture depends on the effects of soil microbial activities and soil fertility. To improve soil health for the upland crops, this study evaluated a relationship between soil chemical properties and soil microbial diversities at 25 sites in upland soils in Gyeongnam Province. The average nutrients in the upland soils were 1.7 times for available phosphorous, 1.4 times for exchangeable potassium and 1.5 times for exchangeable calcium higher compared to recommend concentrations in the upland soils. We found a significant positive correlation between the soil organic matter and the soil microbial biomass C (p<0.01). Contents of organic matter and dehydrogenase in the inclined piedmont soils were significantly higher than those in the other topographical soils (p<0.05). In addition, concentrations of organic matter and microbial biomass C in the loam soils were significantly higher than in the silt loam soils (p<0.05). In principal component analyses of chemical properties and microbial populations in the upland soils, our findings suggested that available phosphorous should be considered as potential factor responsible for the clear upland soils differentiation. The soil organic matter was positive correlation with Bacillus sp. and fungi, whereas soil pH was also positive correlation with Pseudomonas sp. in upland soils.