• Title/Summary/Keyword: Korean native strain

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Purification and Enzymatic Characteristics of the Bacillus pasteurii Urease Expressed in Escherichia coli (Escherichia coli에서 발현된 Recombinant Bacillus pasteurii Urease의 정제 및 효소학적 특성)

  • 이은탁;김상달
    • Microbiology and Biotechnology Letters
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    • v.20 no.5
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    • pp.519-526
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    • 1992
  • The gene coding for urease of alkalophilic Bacillus pasteurii had been cloned in Escherichia coli previously. The urease protein was purified 63.1-fold by TEAE-cellulose, DEAE-Sephadex A-50, Sephadex G-150 and Sephadex G-200 chromatographies with a 7.3% yield from the sonicated fluid of the E. coli HB1Ol(pBUll) encoding B. pasteurii urease gene. The ureases of E. coli (pBUll) and B. pasteurii possessed as a $K_m$ for urea, 42.1 mM and 40.4 mM, respectively. They hydrolyzed urea with $V_{max}$ of 86.9$\mu$mol/min and 160$\mu$mol/min, respectively. Both ureases were composed with four subunits (Mrs 67,000) and a subunit (Mr 20,000). The molecular weight of both native enzymes was Mr 280,OOO$pm$10,000 determined by gel filtration chromatography and Coomassie blue staining of the subunits. The optimal reaction pH of both ureases were pH 7.5. The ureases were stabled in pH 5.5-10.5. The optimal reaction temperature of both ureases were $60^{\circ}C$, and the ureases were stable for an hour at $50^{\circ}C$, 40min at $60^{\circ}C$ and 10 min at $70^{\circ}C$ The activity of both enzymes were inhibited completely by $Ag^{2+}$, $Hg^{2+}$, $Zn^{2+}$, $Cu^{2+}$, and were inhibited 60% by CoH, 30% by $Fe^{2+}$ and 10% by $Pb^{2+}$. However it was increased by the addition of $Sn^{2+}$, $Mn^{2+}$, $Mg^{2+}$ at concentration of $1{\times}10^{-3}$M. Both ureases were inhibited completely by p-CMB and acetohydroxamic acid. The urease expressed in E. coli (pBU11) was inhibited 70% by SDS. The urease of B. pasteurii was inhibited 40% by hydroxyurea, whereas the recombinant urease of E. coli strain was inhibited 17%. Both enzymes were not inhibited by cyclohexanediaminetetraacetic acid (CDTA) and ethylendiaminetetraacetic acid (EDTA).

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Material and Behavior Characteristics of Lightweight Embankment for Road Constructed on Soft Ground (연약지반에 시공된 도로용 경량성토체의 재료 및 거동특성)

  • Yea, Geu-Guwen;Lee, Yong-Jae;Kim, Hong-Yeon;Yoon, Gil-Lim;Han, Sang-Hyun
    • Journal of the Korean Geosynthetics Society
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    • v.17 no.2
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    • pp.41-49
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    • 2018
  • The purpose of this study is to fabricate a full scale road embankment using lightweight air foamed soil as a soil material on soft ground and to investigate its material characteristics and behavior in order to promote dredged soil utilization and minimize ground improvement. As a result of the laboratory test of the onsite mixed samples, the total unit weight of the specimens decreased almost linearly until curing 28 days. In particular, the total unit weight after 28 days of curing was reduced to about 81% of the slurry state before curing, which will be useful in the formulation of similar native soil materials in the future. The unconfined compressive strength began to decrease with the 14th day of curing as shown in the previous study. When the cement content is increased, the strength decreases sharply at a small strain change after the occurrence of the maximum compressive strength, and the maximum strength is exhibited in a range of a smaller axial strain than normal range. The settlement at the surface layer of the ground due to the lightweight embankment was about 1 / 2.75 of the soil embankment and was in agreement with the unit weight ratio (1 / 2.7) of the embankment materials. This indicates the cause and effect of the settlement due to the difference in self weight of the embankments. Also, the difference in settlement between soil and lightweight embankment increased with increasing depth. This shows that the difference in the point at which the settlement is terminated is clear. The ground horizontal displacement under the lightweight embankment was about 15~20% smaller than that of the soil embankment and the depth of occurrence was also 4.5~5.0m shallower in the lightweight embankment.