• Title/Summary/Keyword: 토양 효소 활성

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Expression and Purification of Three Lipases (LipAD1, LipAD2, and LipAD3) and a Lipase Chaperone (LipBD) from Acinetobacter schindleri DYL129 (Acinetobacter schindleri DYL129 유래의 3개 lipases와 chaperone의 발현과 정제)

  • Kim, Sun-Hee;Lee, Yong-Suk;Jeong, Hae-Rin;Pyeon, Hyo-Min;You, Ju-Soon;Choi, Yong-Lark
    • Journal of Life Science
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    • v.29 no.4
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    • pp.492-498
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    • 2019
  • Previously, three kinds of lipases, lipAD1, lipAD2, and lipAD3, and lipase chaperone, lipBD, of Acinetobacter schindleri DYL129 isolated from soil sample were reported. In this report, three lipase and lipase chaperone were cloned into the pET32a(+) or pGEX-6P-1 vectors for protein expression in Escherichia coli, and named as pETLAD1, pETLAD2, pETLAD3 and pETLBD or pGEXLAD1, pGEXLAD 2, pGEXLAD3 and pGEXLBD, respectively. Protein expression rate was higher in pET system than in pGEX system. Although LipAD1 and LipAD2 were produced as inclusion bodies, their expression levels were high. So LipAD1 and LipAD2 could be solubilized in 8 M urea buffer and purified. LipAD3 and LipBD were overexpressed in soluble form and purified. Those proteins were purified by His-tag affinity chromatography connected in AKTA prime system. The activities of the purified lipases were demonstrated with 1% tributyrin agar plate. After purification, molecular mass was determined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis. LipAD1 showed high activity toward ${\rho}$-nitrophenyl acetate and ${\rho}$-nitrophenyl butyrate, LipAD2 showed high activity toward ${\rho}$-nitrophenyl acetate and ${\rho}$-nitrophenyl myristate, and LipAD3 showed high activity toward ${\rho}$-nitrophenyl acetate, ${\rho}$-nitrophenyl butyrate, and ${\rho}$-nitrophenyl miristate, respectively. Three lipases, LipAD1, LipAD2, and LipAD3, showed optimal reaction at $50^{\circ}C$ using ${\rho}$-nitrophenyl butyrate, as substrate.

Ecological Studies on the Transition of Sheath Blight of Rice in Korea (한국(韓國)에서의 벼 잎집무늬마름병 발생변동(發生變動)에 관(關)한 생태학적(生態學的) 연구(硏究))

  • Yu, Seung-hun
    • Korean Journal of Agricultural Science
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    • v.4 no.2
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    • pp.283-316
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    • 1977
  • In an attempt to obtain a basic information to develop an effective integrated system of controlling sheath blight of rice in Korea, the transition of this disease, the variation of cultural characters and pathogenicity of the pathogen, environmental conditions affecting the disease outbreak and varietal resistance have been investigated. 1. Rice sheath blight which has been minor disease in the past was widely spread, especially since 1971. This disease has promptly spread all over the country and infected 65.2% of total rice growing area in 1976. Various factors are considered to be related to such transition of this disease. Above all, increace of application of nitrogenous fertilizer, early season and earlier cultivation of rice, introduction of more susceptible "Tongil" varieties etc. must be important factors influencing the outbreak of this disease. 2. Great variations in cultural characteristics-such as mycelial growth rate, color of the medium, amount of the aerial mycelium, shape and color of the sclerotia- and in the pathogenicity of isolates of the pathogen, Thanatephorus cucumeris Dank were observed. The optimum temperature for mycelial growth also varied with isolates, from $25^{\circ}C$ to $30^{\circ}C$. There were not necessarily any correlation between curtural characteristics and pathogenicity of isolates of Thanatephorus cucumens. 3. Mycelial grow th of isolates of Thanatephorus cucumens on the PDA medium were correlated with the air temperatures of the region where the isolates were collected. The isolates from the regions with high temperature grew well on PDA medium at $35^{\circ}C$ than those from the region with low temperature, on the other hand, the isolates from the regions with the low temperature grew well on the same medium at $12^{\circ}C$ than those from the regions with high temperature. 4. Pectin polygalacturonase (PG) and cellulase (Cx) were most active on the 3rd day after inoculation on the leaves of rice plant with Thanatephorus cucumeris, whereas pectin methylestrase (PE) was most active on the 4th day after inoculation. Relationship between the activities of PE of isolates and the strength of pathogenicity of isolates was obtained, but PG and cellulase activities were not correlated with pathogenicity of isolates. 5. The tolerence of sclerotia from in-vitro culture to low temperature varied with their water content, the dried cultural sclerotia were more tolerent than wet ones, Dried cultural sclerotia maintained almost 100% germinability for 45 days at $-20^{\circ}C$, whereas wet sclerotia lost viability at $-5^{\circ}C$. The germination ratio of the sclerotia after overwintering changed from 18% to 70% according to the water content of the test paddy fields and the ratio was low in wet paddy condition. 6. To investigate the host range of this fungi in and near paddy field, 17 weeds were inoculated with fungi. The lesions of sheath blight disease was obserbed on Sagittaria trifolia L., Echinochloa crusgalli P. Beauv., Monochoria vaginal is Presl, Polygonum Hydropiper L., Eclipta prostrata L., Digitaria sanguinalis Scapoli. 7. When the level of nitrogen applied was doubled over standard level, total nitrogen content in rice sheath increased, ami when silicate was applied, starch content in rice sheath decreased, inducing the rice plants more susceptible to sheath blight disease. Increased dressing of potash ferilizer reduced the incidence of sheat blight disease. 8. The percentage of infected stems in the early period increased more in the narrow hill plot than in the wide hill plot, but in the late period this tendency was inversed; the percentage of infected stems as well as severity in the wide hill plot increased more compared to the narrow hill plot, and the disease severity in the one plant per hill plot was also low. The number of stems in the wide hill plot was more than the number of stems in the narrow hill plot. This indicates that the microclimate, such as the relative humidity, in the narrow hill plot was more favorable for the development of this disease. 9. There was a high negative correlation between the disease severity of varieties to the sheath blight and the maturity of the varieties, that is, the early varieties were more susceptible than the late ones, and much-tillering varieties usually showed more infection than less tillering varieties. 10. No relationship was obtained between the percentage of infected stems in the early period and the severity after heading, whereas a distinct relationship was obtained between former and latter after Aug. 10.

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