• Title/Summary/Keyword: 토양 미생물 효소 활성

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Purification and Some Properties of Rutinosidase from Arthrobacter sp. (Arthrobacter sp.가 생산하는 Rutinosidase의 정제 및 특성)

  • ;Toshio Omori;Tohru Kodama
    • Microbiology and Biotechnology Letters
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    • v.18 no.4
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    • pp.360-367
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    • 1990
  • The several glycoside hydrolysing enzymes related to rutin degradation are found to be rhamnosidase, glucosidase and rutinosidase. Rutinosidase was purified to electrophoretic homogeneity from cell extracts of rutin-degrading strain, MT-57, which was identified as a Arthrobacter sp. Its molecular weight was estimated to be 42, 000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and 40, 000 by gel filtration. The optimum pH for enzyme was found to be 7.5, and relatively stable in alkaline solution. The optimum temperature for enzyme was $45^{\circ}C$, being stable up to $50^{\circ}C$ for 20 min. The Bm value of enzyme for rutin was 0.5 $\mu \textrm m$. The enzyme activity was increased by the chelating agent such as EDTA, $NaN_3$, and 8-hydroxyquinoline, was strongly inhibited by $CO_{2+}, Ni^{2+}$, and $Cu^{2+}$. The enzyme had high substrate specificity in the rutinoside.

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Fermentation Patterns of Chungkookjang and Kanjang by Bacillus licheniformis B1 (Bacillus licheniformis B1에 의한 청국장 및 간장 발효)

  • Lee, Jae-Jung;Lee, Dong-Seok;Kim, Han-Bok
    • Korean Journal of Microbiology
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    • v.35 no.4
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    • pp.296-301
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    • 1999
  • A Bacillus strain from Korean soil was isolated and identified to be Bacillus licheniformis B1 through various biochemical tests, VITEK, and MIDI system analysis. The strain produced extracellular amylase and protease. Whether or not the strain can perform Chungkookjang fermentation with autoclaved soybean and Kanjang fermentation was determined in this study. In Chungkookjang fermentation, browining materials of strong anti-oxidant increased 8-fold, and 2-fold in Kanjang, compared with initiation material for fermentation. Maximal protease activity in Chungkookjang was observed one day after inoculation. Protease activities in Kanjang decreased to the half, and then maintained constant values during fermentation, probably due to the inhibitory effect of salt on protease activities. High molecular mass of nucleic acids was identified in Chungkookjang and Kanjang. Since the nucleic acids were not observed in autoclaved soybean, they seem to be originated from B. licheniformis B1. This study demonstrated successive fermentation of Chungkookjang and Kanjang by B. licheniformis B1 isolated from nature, and suggested possible development of food rich in browing and nucleic acids.

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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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Restriction Mapping of Cloned Pullulanase Gene and Property of Pullulanase Produced in Escherichia coli (pYKL451) and Klebsiella pneumoniae NFB-320 (Klebsiella pneumoniae NFB-320의 Pullulanase 유전자의 제한효소 분석과 효소학적 특성)

  • Yu, Ju-Hyun;Chung, Kun-Sub;Kong, In-Su;Lee, Jung-Kee
    • Microbiology and Biotechnology Letters
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    • v.15 no.6
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    • pp.436-440
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    • 1987
  • Pullulanase gene (pul) of Klebsiella pneumoniae NFB-320 which was cloned previously in Escherichia coli with plasmid pBR322. The gene was analyzed with various restriction enzymes. The cloned gene was contained within n 10 kb BamHI DNA fragment. We constructed the restriction map of the hybrid plasmid pYKL451. The optimum temperatures for pullulanases produced in E. coli (pYKL451) and K. pneumoniae NFB-320 were almost the same, 50-55 $^{\circ}C$. The optimum pHs for the reaction of the enzymes produced by E. coli (pYKL451) and K. pneumoniae NFB-320 was 6.0. Both enzyme preparations were stable under the range of pH 5.0 to 10.0 when those were kept at 40 $^{\circ}C$ for 90 min and were stable until 40 $^{\circ}C$ when allowed to stand for 1hr at various temperatures.

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Effect of Different Fertilization Management Practices on Soil Microbial Activities and Community Structure in Volcanic Ash Citrus Orchard Soil (화산회토 감귤원 토양의 시비관리가 토양미생물활성 및 군집구조에 미치는 영향)

  • Joa, Jae-Ho;Han, Seung-Gap;Won, Hang-Yeon;Lim, Han-Cheol;Hyun, Hae-Nam;Suh, Jang-Sun
    • Korean Journal of Soil Science and Fertilizer
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    • v.42 no.3
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    • pp.222-229
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    • 2009
  • This study was performed to evaluate effect of different fertilization management practices on soil microbial activities and community structure using soil enzyme activities and PLFA contents in volcanic ash citrus orchard soil. Six experiment plots have differently managed based on the recommended application rate(NPK) of chemical fertilizer and compost for 13 years. Experiment plots were composed of no-fertilization(control), compost only, half amount of NPK with compost (1/2NPK+COM), NPK, NPK with compost(NPK+COM), and 3 times amount of NPK(3NPK). Soil samples collected in early March, May, July, and September 2007. Urease activity was high at NPK+COM in March, May, and September. It was higher in NPK+COM than in NPK. Urease activity decreased according to the order NPK>compost>control in March and May; compost>NPK>control in July and September. Dehydrogenase activity was significantly higher in 1/2NPK+COM($4.3ug\;TPF\;g^{-1}\;24h^{-1}$) than in control($2.4ug\;TPF\;g^{-1}\;24h^{-1}$), May. $\beta$-glucosidase activity was significantly higher in NPK and 1/2NPK+COM than in control, May. In March, Total PLFA contents were higher in NPK+COM($349.2n\;mol\;g^{-1}$) than in 3NPK($228.5n\;mol\;g^{-1}$). And that were higher in 1/2NPK+COM($237.8n\;mol\;g^{-1}$) than in 3NPK($133.1n\;mol\;g^{-1}$), May. Distribution ratio of soil microbial groups by PLFA biomaker were not significantly difference in between seasonal and treatments. Principal component analysis by PLFA profiles showed that microbial community in compost and 3NPK plot were different compared with other treatments in March. But Differences in compost and 3NPK plot were not found in May. Our result showed that the change of microbial community structure affected by fertilization effect and seasonable variation.

Screening of Antagonistic Bacteria Having Antifungal Activity against Brown Spot and Sheath Rot of Rice (벼 깨씨무늬병 및 잎집썩음병에 항진균 활성을 갖는 길항 미생물의 탐색)

  • Ryu, Myeong Seon;Yang, Hee-Jong;Jeong, Su-Ji;Seo, Ji-Won;Jeong, Do-Youn
    • The Korean Journal of Mycology
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    • v.47 no.3
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    • pp.259-269
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    • 2019
  • Brown spot and sheath rot of rice are caused by fungal pathogens such as Curvularia lunata, Cochliobolus miyabeanus, and Sarocladium oryzae, and cause losses such as reduced rice yield and quality, which is an enormous problem with serious long-term effects. To search biological control agents of phytopathogenic fungi, five kinds of useful Bacillus-like isolates which are excellent in extracellular enzyme activity and produce siderophore were selected from paddy soil of Sunchang in Korea. The selected isolates were tested for excellent antifungal activity against three of the phytopathogenic fungi that frequently occur in rice, and JSRB 177 strain had the most excellent antifungal activity. Based on the experimental results, JSRB 177 is finally selected as a candidate for biological control and identified to Bacillus subtilis through 16S rRNA sequence analysis. In addition, physiological characteristics of JSRB 177 confirmed by analysis of carbohydrate fermentation patterns and enzyme production ability. Based on the above results, JSRB 177 is expected to be used as a biological control agent for the rice pathogenic fungi. In the future, further studies related to industrialization such as port test and establishment of mass production process are needed.

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.

Studies on the Naringinase Produced by Aspergillus nidulans (Part I) Screening of Naringinase Produced by Fungi and Their Cultural Condition (Aspergillus nidulans가 생산하는 Naringinase에 관한 연구 (제1보) 생산균주의 선정 및 Naringinase의 생산조건)

  • 유주현;문순옥;변유량
    • Microbiology and Biotechnology Letters
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    • v.6 no.2
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    • pp.59-63
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    • 1978
  • The Screening of fungi producing naringinase was done. A strain of Aspergillus midulans showed the highest naringinase activity among 447 strains those were isolated from soil, spoiled citrus friuts and stock cultures. The cultural Conditions of Asp. nidulans for production of naringinase were studied. A strain of Asp. nidulans showed higher activity when it was cultivated at 30$^{\circ}C$ for 3 days on wheat bran media supplemented with 2.0% naringin, 0.2% (NH$_4$)$_2$ SO$_4$ and 0.2% CaCO$_3$.

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Purification and Characterization of Phospholipase D from Actionmycetes KF923 (방선균 KF923이 생산하는 Phospholipase D의 정제 및 특성)

  • 곽보연;윤석후;김창진;손동화
    • Microbiology and Biotechnology Letters
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    • v.32 no.1
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    • pp.78-83
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    • 2004
  • In order to screen microorganisms producing phopholipase D (PLD) had high transphosphatidylation activity, about 1,000 Actinomycetes strains were isolated from the 63 soil samples, collected over 6 local area in Korea. When the hydrolytic activity in the supernatant was determined, 131 strains produced PLD more than 0.3U/$m\ell$. Among 131 culture broths tested, 23 ones had transphosphatidylation activity higher than 20% and finally one strain (Actinomycetes KF923), which had highest hydrolytic and transphophadylation activity, was selected. Actinomycetes KF923 showed the highest hydrolytic activity (13U/$m\ell$) and phosphatidylation activity (95%) after 48 h fermentation using the P medium (yeast extract 1%, peptone 1%, glucose 1.5%, glycerol 1%, $CaCO_3$ 0.4%, pH 7.2). PLD was purified from the culture broth of Actinomycetes KF923 and the specific activity of purified PLD was 567U/mg. The molecular weight of PLD was about 55kD and the optimum pH and temperature were pH 6.0 and $60^{\circ}C$, respectively. The stability of PLD toward pH and temperature were high around pH 8.0 and below $40^{\circ}C$ Special metal ions were not necessary to the PLD activity.

Purification and Characterization of Phospholipase D from Actinomycetes KF923. (방선균 KE923이 생산하는 Phospholipase D의 정제 및 특성)

  • 곽보연;윤석후;김창진;손동화
    • Microbiology and Biotechnology Letters
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    • v.31 no.4
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    • pp.389-394
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    • 2003
  • In order to screen microorganisms producing phopholipase D (PLD) had high transphosphatidylation activity, about 1,000 Actinomycetes strains were isolated from the 63 soil samples, collected over 6 local area in Korea. When the hydrolytic activity in the supernatant was determined, 131 strains produced PLD more than 0.3 U/ml. Among 131 culture broths tested, 23 ones had transphosphatidylation activity higher than 20% and finally one strain (Actinomycetes KF 923), which had highest hydrolytic and transphophadylation activity, was selected. Actinomycetes KF923 showed the highest hydrolytic activity (13 U/ml) and phosphatidylation activity (95%) after 48 h fermentation using the P medium (yeast extract 1%, peptone 1%, glucose 1.5%, glycerol 1%, $CaCo_3$ 0.4%, pH 7.2). PLD was purified from the culture broth of Actinomycetes KF923 and the specific activity of purified PLD was 567 U/mg. The molecular weight of PLD was about 55 kD and the optimum pH and temperature were 6.0 and $60^{\circ}C$, respectively. The stability of PLD toward pH and temperature were high around pH 8.0 and below $40^{\circ}C$. Special metal ions were not necessary to the PLD activity.